Application of Enzyme-Assisted Extraction on the brown seaweed Fucus vesiculosus Linnaeus (Ochrophyta, Fucaceae) to produce extracts for a sustainable agriculture.

Application of Enzyme-Assisted Extraction on the brown seaweed Fucus vesiculosus Linnaeus (Ochrophyta, Fucaceae) to produce extracts for a sustainable agriculture. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Application of Enzyme-Assisted Extraction on the brown seaweed Fucus vesiculosus Linnaeus (Ochrophyta, Fucaceae) to produce extracts for a sustainable agriculture. Manon Choulot, Chirelle Jabbour, Anne-Sophie Burlot, Lun Jing, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5331195/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract The agricultural sector plays a crucial role in sustaining population growth and ensuring our well-being. However, as this sector faces numerous challenges due to environmental conditions and the increasing population, it is essential to identify natural alternatives that promote sustainable farming practices and protect the environment. Seaweed-based extracts have gained popularity in agriculture because of their numerous benefits for plant growth and health. This study focuses on Fucus vesiculosus Linnaeus, a prevalent brown seaweed species found along the Brittany coast. The research involved comparing Enzyme-Assisted Extraction (EAE) with conventional aqueous extraction methods for the purpose of determining the effectiveness of EAE in producing enriched extracts that could have an interesting agricultural application. The results indicates that enzymatic extraction of Fucus vesiculosus significantly increased the content of neutral sugars by 34% and reducing sugars by 21% in the extracts, compared to conventional aqueous extraction (WE). Regarding Plant Growth Regulators, the levels of Isopentenyladenosine (iPR) and Cis zeatin (cZ) were enhanced by 6 times and 28 times, respectively, when using EAE instead of WE. Additionally, the total phenolic content was notably higher in EAE extracts, showing a twofold increase over WE extracts. Furthermore, the various extracts demonstrated superior antioxidant activity compared to raw Fucus vesiculosus powder. Thus, this study confirms that EAE is an effective method for enriching Fucus vesiculosus extracts with various compounds that can play a vital role in agriculture. Brown seaweed Fucus vesiculosus Enzyme-Assisted Extraction Biochemical Composition Elements Plant Growth Regulators Figures Figure 1 Figure 2 1. Introduction Over the last few years, the agricultural sector is facing numerous challenges due to a burgeoning global population, climate warming, severe weather events, the over use of mineral fertilizers and in the same time the necessity to produce food in more efficient and sustainable ways. This context led again to a broader interest in natural alternatives to preserve environment and produce quality food. One of the most popular, intensively developing groups of biostimulants are seaweed extracts. Biostimulants are the most promising short-term markets for seaweed, projected to reach USD 4.4 billion by 2030. According to Global Seaweed New and Emerging Markets Report (2023), seaweed-based biostimulants can expect to see significant growth over the next few years as additional investment goes into research and development for improving efficacy, and as more and more seaweed processors are exploiting seaweed biomass as a whole in a biorefinery approach to create multiple added-value products. For centuries, coastal populations in various regions across the globe have harvested a diverse range of seaweeds, source of livelihood and survival (Jacob et al. 2012 ). These seaweeds were first used for domestic purposes, such as human health, consumption, or glass production (Arzel 1986). Seaweeds have also been employed as fertilizers, soil enhancers, and for soil pH adjustment. In regions such as Brittany and along the French Atlantic coast, seaweed was traditionally collected by coastal farmers using large rakes following storms and directly applied to fields. This traditional practice persists on the island of Batz in northern Brittany and the Rhuys peninsula in southern Brittany in France (Choulot et al. 2022 ). The application of seaweed-based products in agriculture has been gaining momentum in crop production systems over the past 30 years. Today, seaweed used for biostimulants is mainly brown wild-harvested especially Durvillaea potatorum and Ecklonia maxima (Laminariales) or Ascophyllum nodosum (Fucales) (Rousseau et al. 2023 ). Various liquid products are present on the market such as Seasol with Durvillaea potatorum , Kelpak with Ecklonia maxima , or Fertileader, AlgaGreen and Maxicrop with Ascophyllum nodosum (Choulot et al. 2022 ). Rich in wide variety of compounds – macronutrients and micronutrients, alginates and oligosaccharides, Plant Growth Regulators (PGRs), vitamins, and osmolytes – seaweeds extracts are used as agricultural input products in different forms, such as biofertilizers, liming materials, soil improvers, plant biostimulants, and fertilizing product blends (e.g., Bikovens et al. 2017 ; Michalak and Baśladyńska, 2021 ; Mzibra et al. 2021 ; Choulot et al. 2022 ; Krautforst et al. 2023 ; Spain et al. 2024 ). Among the harvested algae in France, Fucus vesiculosus Linnaeus, commonly known as bladder wrack, is most commonly found on sheltered rocky shores subject to some degree of disturbance, such as scour (Bunker et al. 2017 ). In Brittany, F. vesiculosus is particularly abundant, forming extensive intertidal zones characteristic of the mid and low-mid intertidal zones. This species can grow to sizes ranging from tens centimeters to one meter in length, with a lifespan typically between 2 and 5 years (Knight and Parke 1950 ). The biomass of Fucus can now reach several hundred to several thousand fresh tons per year (Mesnildrey et al. 2012 ; CSAVM 2023, personal communication), continuing to play a significant role in local economies for various industries such as cosmetics and agriculture. In agriculture, due to its unique chemical composition, Fucus sp. biomass is being tested for the use in agriculture as a soil additive, positively influencing plant growth, for example: oat (Bikovens et al. 2017 ), garden cress (Michalak and Baśladyńska, 2021 ) or sorghum (Krautforst et al. 2023 ). For the production of seaweed based biostimulants, alkaline extraction using potassium hydroxide is currently the most common method, constituting approximately 80% of all biostimulant production (Choulot et al. 2022 ; Global Seaweed New and Emerging Markets Report 2023). However, some bioactive compounds such as PGRs, vitamins, and antioxidants are susceptible to degradation under harsh extraction conditions (extreme pH or/and high temperatures) (Sharma et al. 2014 ). The complexity of the cell wall, with strong interactions between fibrillar, matrix polysaccharides, polyphenols, and proteins, hampers the release of compounds during extraction. Numerous physicochemical and biological methods have been developed to recover active compounds from algal biomass (Kadam et al. 2013 ; Michalak and Chojnacka, 2014 ). In the context of climate change and socio-environmental concerns, the blue circular economy, selecting an appropriate extraction method must balance metabolites selectivity and recovery, cost-effectiveness, and eco-friendly processes based on green and sustainable chemistry and engineering. Currently, Enzyme-Assisted Extraction (EAE), Microwaves Assisted Extraction (MAE) or Ultrasound-Assisted Extraction (UAE) are among the techniques most mentioned in the literature for natural biomass processing, used alone or combined (Wijesinghe and Jeon 2012 ; Michalak and Chojnacka, 2014 ; Choulot et al. 2022 ; Spain et al. 2022; Le Guillard et al. 2023 ; Krautforst et al. 2023 ; Choulot et al. 2023 ). The processes are described as feasible and efficient alternatives to traditional extraction methods and might be transposed on a large scale for seaweed processing in some industrial sectors. Due to the highly site-specific activity of the hydrolytic enzymes, enzymatic hydrolysis process has been an excellent, environmentally-friendly method to produce some active compounds. EAE effectively extracts valuable components from seaweed without the need for denaturing conditions such as solvents or high temperatures. The effectiveness of EAE in seaweed liquefaction has been demonstrated through the use of two types of enzymes: proteases and carbohydrases. Research has consistently shown that the use of enzymes can significantly enhance the value of seaweed extracts or compounds (Hardouin et al. 2014 ; Michalak and Chojnacka, 2014 ; Spain et al. 2022; Choulot et al. 2023 ). EAE is currently used in various industrial applications, including energy (methanation), agriculture, feed, and food (Michalak and Chojnacka, 2014 ; Wijesinghe and Jeon, 2012 ; Terme et al., 2020; Choulot et al. 2023 ). Several works explore EAE for the production of molecules of interest from seaweeds (Hardouin et al. 2014 ; Obluchinskaya et al. 2021; Getachew et al. 2022; Spain et al. 2022; Pliego-Cortes et al. 2023) but no work reports the use of this technique for the production of biostimulants with Fucus vesiculosus biomass. This study explores the potential benefits of employing Enzyme-Assisted Extraction on Fucus vesiculosus Linnaeus ( Ochrophyta , Fucaceae ) by analyzing specific compounds pertinent to agriculture, including Plant Growth Regulators, polyphenols and macro/micronutrients. 2. Material and methods Fucus vesiculosus Linnaeus powder (< 904 µm) was purchased from Groupe Roullier (Saint-Malo, France). Seaweeds were collected in Brittany (France) in July 2019 and dried in an industrial drying tunnel at 100 ± 20°C. Right after drying, F. vesiculosus was ground and stored in opaque plastic buckets at room temperature for further experiments. 2.1 Enzyme-Assisted Extraction (EAE) and Water Extraction (WE) of Fucus vesiculosus Linnaeus A proximate analysis was conducted to aid selection of appropriate enzymes for seaweed hydrolysis. Seventeen commercial enzymatic preparations were separately evaluated for the extraction of Fucus vesiculosus . The operational pH and temperature were chosen according to the suppliers’ recommendations (data not shown). Three commercial enzymatic preparations were thus selected – two glycosidases, G1 and G3, and one protease, P2 (Table 1 ) with respective activities based on the supplier's datasheets. Table 1 Characteristics of the enzymatic preparations used for extraction of Fucus vesiculosus. Enzyme Activity Source Form Operational pH Operational temperature Glycosidase G1 β-glucosidase Aspergillus niger Powder 4 50–65°C G3 β-1,3-glucanase and Botrytis glucanase Trichoderma longibrachiatum Liquid 5 50°C Protease P2 Protease (neutral) (endo) Bacillus amyloliquefaciens Powder 7 50°C EAE was performed as described by Hardouin et al. ( 2014 ), with minor modifications. The seaweed powder (2.5% solid/liquid ratio (w/w)) was added to 1 L of distilled water in a jacketed glass reactor vessel. The pH was adjusted for each enzyme using either hydrochloric acid (HCl) or sodium hydroxide (NaOH). Enzymes added at a concentration of 5% (enzyme/seaweed ratio (w/w)) were placed in a 50°C water bath with 150 rpm of continuous tangential agitation. After 17 h, the enzymes were denatured by raising the temperature to 90°C for 15 min. Water extracts (WE) were obtained from seaweed powder processed in the same conditions (including the denaturation step) without enzyme addition. The extracts (WE and EAE) were centrifuged (5000 g for 15 min at 20°C) to remove the undigested residue, and then freeze-dried. Each extraction, WE and EAE, was performed in triplicate. The dry matter extraction yield (EY) was obtained by calculating the ratio between the total weight of dry extract (m 1 ) and the total weight of dry matter in the reactor (m 2 ), and results are expressed in percentage (Eq. 1 ). $$\:EY=\:\frac{m1\left(g\right)}{m2\:\left(g\right)}\:x\:100\%\:$$ 1 2.2 Analysis of biochemical composition Analyses were carried out on seaweed powder and on extracts (EAE and WE). Firstly, for some analyzes (total neutral sugars and proteins content), the F. vesiculosus powder was subjected to acid hydrolysis to improve their recovery and quantification according to Hardouin et al. ( 2014 ). Thus, 5 mL of 1 M HCl were added to 10 mg of dry seaweed and incubated for 2 h at 100°C in a dry bath. Then, the solution was neutralized by adding 5 mL of 1 M NaOH. All chemical analyses were performed in triplicate (N = 3). Secondly, for other analyses (including total phenolic content and antioxidant activity), a multistep extraction process was conducted on F. vesiculosus powder. Initially, 5 mg of F. vesiculosus powder was mixed with 1 mL of distilled water and incubated for 2 hours at 100°C in a thermoshaker set to 1000 rpm. The resulting supernatant was then collected. The remaining precipitate underwent a second extraction with the addition of 1 mL of 1 M HCl, followed by another 2-hour incubation under the same conditions. The supernatant from this second extraction was combined with the first and neutralized using 1 mL of 1 M NaOH. Subsequently, the precipitate was resuspended in 1 mL of 5 M NaOH and incubated again. The final supernatant was collected and neutralized with 1 mL of 5 M HCl. All chemical analyses were performed in triplicate (n = 3). 2.2.1 Total sugars Analysis of total sugars content was performed according to the method described by Dubois et al. ( 1956 ). 1 mL of tested sample or standard (glucose) (Avocado Research Chemicals Limited) was inserted into a hemolysis tube. 15 µL of 75% phenol (Prolabo) were added. After stirring the solution, 2.5 mL of 96% H 2 SO 4 were added to the tube. The solution was cooled in an ice bath for 10 min and then placed in a water bath at 30°C for 10 min. After stirring, the solutions were ultrasonicated for 5 s to remove micro air bubbles. The absorbance was read at 490 nm with a spectrophotometer UV-1800 (Shimadzu) and the results are expressed in percentage of dry seaweed (d.s.) or percentage of dry extract (d.e.). The standard glucose range was performed at 20, 40, 60, 80 and 100 µg mL − 1 . 2.2.2 Reducing sugars This analysis was carried out on extracts (WE and EAE) according to the Miller method (1959). First, a solution of 3,5-dinitrosalicylic acid (DNS) was prepared by dissolving 1 g of DNS in 70 mL of 0.8 M NaOH while heating. After that, 30 g of sodium potassium tartrate were added before making up the solution to 100 mL with distilled water, which was then kept in the dark. 500 µL of the sample or standard solution (glucose) (Avocado Research Chemicals Limited) were added to 500 µL DNS reagent in Eppendorf tubes. The sample solutions were heated in a water bath for 10 min at 95°C and cooled immediately in an ice bath for 5 min. Then, the samples were incubated 15 min in the dark before taking a 200 µL aliquot and measuring its absorbance at 540 nm on a 96-well plate using a Multiskan GO plate reader (Thermo Scientific). The results are expressed in percentage of dry mass of extract (d.e.). The glucose concentrations were determined using a range of standard solutions at 50, 100, 150, 200, 250, 300, 400 and 500 µg mL − 1 . 2.2.3 Uronic acids The composition of uronic acids was analyzed using the colorimetric method originally developed by Blumenkrantz and Asboe-Hansen in 1973, and later modified by Filisetti-Cozzy and Carpita in 1991. When exposed to hot concentrated strong acid, uronic acids dehydrate and then cyclize, forming derivatives of 5-formylfuroic acid. These derivatives react with meta-hydroxydiphenyl (MHDP) to produce a pink chromophore that absorbs light at λ = 525 nm. In addition, the coloration is brighter in the presence of borate, but remains very sensitive to interference from neutral sugars, which stain in the presence of concentrated sulphuric acid. For this reason, potassium sulfamate is added to limit these interferences (Pierre 2010 ). Two hundred µL of the tested sample or standard (glucuronic acid from Acros Organics) were placed into a hemolysis tube. Subsequently, twenty µL of 4 M sulfamic acid (Alfa Aesar) were added. After stirring the mixture, 1.2 mL of a 75 M sodium tetraborate solution (from Merck) was carefully added. The tubes were then capped and incubated at 80°C for 20 minutes. Following incubation, the tubes were cooled in ice for 5 minutes. Forty µL of a 0.15% aqueous solution of meta-hydroxydiphenyl (MHDP) (Acros Organics) were then added. The reaction was allowed to proceed for 10 minutes before measuring the optical density at 525 nm using a Shimadzu UV-1800 spectrophotometer. The glucuronic acid standard curve was prepared with concentrations of 20, 40, 60, 80, and 100 µg mL − 1 . The results were expressed as mg/g of dry mass of seaweed (d.s.) or as a percentage of dry mass of extract (d.e.). The concentration and content of uronic acids were calculated in the same manner as for total sugars. 2.2.4 Proteins The total amount of proteins was determined using the colorimetric method developed by Smith et al. ( 1985 ). 25 µL of sample or standard solution were placed in a 96-well flat-bottom microplate, to which was added 200 µL of the kit reagent (Pierce BCA Protein Assay Kit, Thermo Scientific). The microplate was incubated at 37°C for 30 min before reading at 562 nm on a Thermo Scientific™ Multiskan™ GO Microplate Spectrophotometer. Bovine Serum Albumin (BSA) was used as the standard, with concentrations of 50, 100, 250, 500, 1000 µg mL − 1 . The results were expressed in percentage of dry mass of seaweed (d.s.) or percentage of dry mass of extract (d.e.). 2.2.5 Ash The mineral matter content was determined for F. vesiculosus powder according to the method of Hardouin et al. ( 2014 ). The sample was placed in a crucible and calcined by ignition. The ash content was determined thermogravimetrically after calcination of 100 mg of seaweed powder followed by a passage for 2 h in a Carbolite CSF Muffle Furnace (UK) at 585°C. The final mass corresponds to the mineral matter in the sample and was expressed in percentage of dry mass of seaweed (d.s.). 2.2.6 Multi-elemental composition The total contents of elements in F. vesiculosus and extracts (WE and EAE) were determined using Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) (Agilent bench-top simultaneous optical emission Ar-ICP spectrometer (model 720)) following microwave-assisted closed-vessel digestion (Multiwave PRO microwave reaction system (Anton Paar GmbH)). For this purpose, 0.25 g of sample was placed in a PTFE (Polytetrafluoroethylene) vessel, to which were added 5 mL of 65% HNO 3 (Merck KGaA, Darmstadt). Then, the mixture underwent digestion for 60 min at a maximum temperature of 190°C. After cooling at room temperature, samples were adjusted to 25 g with deionized water and placed in a cool chamber until measurement. Analysis of nitrogen was realized using an elemental FLASH 2000 CHNS analyzer (Thermo Fisher Scientific, Waltham, MA, United States). Briefly, 2.5 mg of the sample were put in an Universal Soft Tin Containers (OD: 5 mm; H = 8 mm, V = 157 µL from Thermo Fisher Scientific, Waltham, MA, United States). Calibration was released using a standard of aspartic acid (Merck, Darmstadt, Germany). Analysis was conducted with the instruction of the manufacturer. The results were expressed as mg kg − 1 of dry mass of seaweed (d.s.) or mg kg − 1 of dry mass of extract (d.e.). Analyses were performed in duplicate. 2.2.7 Plant Growth Regulators (PGRs) The analysis of Plant Growth Regulators was performed on F. vesiculosus raw material without acidic pretreatment, and extracts (EAE and WE). Methods for the quantification of Plant Growth Regulators have been previously described (Haddad et al. 2018 ; Réthoré et al. 2021 ). 20 mg of sample were suspended in H 2 O (Milli-Q, 18.2 MΩ·cm, Millipore) containing isotope-labelled internal standards. The extracts were then purified by SPE (Solid phase extraction) using Evolute express CX (cation exchange), Biotage express plates and ABN columns (Acidic, Basic and Neutral) for cytokinins and other PGRs according to the manufacturer’s protocol. The analyses of PGRs were performed by LC-MS using a Nexera X2 UHPLC system (Shimadzu) coupled to a QTrap 6500 + mass spectrometer (Sciex). The separation was carried out by injecting a 2 µL sample into a Kinetex Evo C18 core–shell column (100 × 2.1 mm, 2.6 µm, Phenomenex) at a flow rate of 0.7 mL min − 1 , with a column temperature maintained at 40°C. The mobile phases were composed of solvent A Milli-Q water containing 0.1% formic acid, and solvent B acetonitrile LCMS grade containing 0.1% formic acid. The analyses were performed in scheduled MRM mode. An external calibration curve was established for each PGR. A quality control sample containing all external and internal standards was injected every 10 samples to assess the system stability. The calculated concentration was corrected by the internal standard recovery rate. The results were expressed in µg kg − 1 of dry mass of seaweed (d.s.) or dry mass of extract (d.e.). 2.2.8 Total phenolic content (TPC) Folin-Ciocalteu method is commonly used for measuring the total phenolic content (TPC). This assay is based on the reduction-oxidation (redox) reactions, which are usually considered to be relatively stoichiometric and on the redox potential of the phenolic hydroxyl group (Hagerman and Butler 1989 ). This analysis was performed directly on F. vesiculosus raw material, on extracts after EAE and on WE from F. vesiculosus . Briefly, in a 96-well microplate, 20 µL of samples (1 mg mL -1 ) or standard (Phloroglucinol from Sigma Aldrich) was added to 100 µL of Folin-Ciocalteu reagent (2N) followed by an incubation of 5 min at 30°C. Then, 80 µL of Na 2 CO 3 solution (7.5%; m/v) was added. The microplate was incubated in the dark for 30 mins min at 40°C. Optical density was measured at 760 nm (Waterhouse 2003; Singleton 1999). Phloroglucinol was used as standard prepared in different concentrations ranging from 0 to 1000 µg mL -1 . The results were expressed in milligrams of Phloroglucinol Equivalent (PE) per g of dry mass of seaweed (d.s.) or dry mass of extract (d.e.). 2.3 Evaluation of antioxidant activities - DPPH Free radical scavenging assay DPPH with stable groups is a typical reagent for evaluating antioxidant activity and can be used as a representative to appraise the scavenging activity of antioxidant compounds against free radicals. This analysis is based on modified method of Guo and Liu (2012). A series of ascorbic acid solutions, served as control, was prepared in different concentrations (0–30 µg mL -1 ). 100 µL of 0.25 mM DPPH solution were introduced to 100 µL sample solution or control on a 96-well microplate. Sample solutions from EAE and WE extracts were prepared in different concentrations by diluting the stock solution in methanol (0-1000 µg mL -1 ). Before reading the optical density at 517 nm, all samples were incubated at 40°C for 30 min in the dark. Percentage of inhibition was calculated using the following formula (2): $$\:I\:\left(\%\right)=\:{\left[\right(A}_{C}-{A}_{S})/{A}_{C}]\:x\:100$$ 2 where: I (%) = inhibition (expressed in %), A C = absorbance of control, and A S = absorbance of samples. IC 50 , corresponding to the concentration sufficient to obtain 50% of a maximum scavenging capacity of samples was determined based on the regression obtained from dose–response curve. 2.4 Statistical analyses The statistical analyses were carried out using R software and RStudio interface, 2023.09.01 version, to evaluate the statistically significant differences (for p < 0.05) between examined groups. Multiple comparison analyses were used for this comparative study between extracts obtained from F. vesiculosus . They include parametric (ANOVA) and non-parametric (Kruskall-Wallis) tests after checking the normality and equal variances of data. These analyses used various packages such as rstatix, RcmdrMisc, multcomp, rcompanion and FSA (Fox et al. 2024 ; Hollander et al. 1973 ; Chambers et al. 1992). 3. Results 3.1 Fucus vesiculosus powder 3.1.1 Proximate composition Figure 1 presents the biochemical composition of the F. vesiculosus raw material. Although Fucus vesiculosus is a very common seaweed species in the Brittany coast, little research has been conducted on the study of its biochemical composition. According to the results, F. vesiculosus is mainly composed of neutral sugars which constitute 22% of d.s. Proteins are the second important component found in F. vesiculosus which constitute 17% of d.s. The uronic acids constitute 12% of d.s. however there is no information was found in the literature concerning the content of uronic acids in Fucus vesiculosus biomass. In addition, the raw F. vesiculosus presents 39.8 ± 9.8 mg eq PE g − 1 d.s. of total phenolic content which consists of approximately 4% of dry weight. 3.1.2 Multi-element analysis Multi-element composition of Fucus vesiculosus is presented in Table 2 . Elements are classified as macroelements (essential for plant growth and development) and microelements (necessary for a number of physiological functions). The three main elements, in term of concentration, present in F. vesiculosus are potassium and sodium with 25 993 mg Kg − 1 d.s. and 25 497mg Kg − 1 d.s., respectively and calcium with 15 966 mg Kg − 1 d.s. Table 2 Multi-element composition of Fucus vesiculosus (mg kg − 1 d.s.) (mean ± SD; n = 2) Element F. vesiculosus powder material Macroelements Potassium 25 993 ± 312 Sodium 25 497 ± 312 Calcium 15 966 ± 335 Sulphur 10 818 ± 834 Nitrogen 8 831 ± 762 Magnesium 8 603 ± 155 Phosphorus 716 ± 9 Microelements Iron 569 ± 11 Boron 118 ± 3 Manganese 106 ± 2 Zinc 25 ± 1 Copper 2 ± 0 3.1.3 Plant Growth Regulators Table 3 represents the content of Plant Growth Regulators in in Fucus vesiculosus . Table 3 Content of Plant Growth Regulators in Fucus vesiculosus (µg kg − 1 d.s.; mean ± SD; n = 3). Plant Growth Regulators Content in F. vesiculosus Salicylic acid (SA) 113 ± 3 Auxin Indole-3-acetic acid (IAA) 54 ± 1 1-amino-1-cyclopropane-carboxylic acid (ACC) 37 ± 3 Dihydrojasmonic acid (DJA) 8 ± 0 Abscisic acid (ABA) 2 ± 0 Cytokinins Cis zeatin (cZ) < 0.1 Isopentenyladenine (iP) 1 ± 0 Isopentenyladenosine (iPR) 1 ± 0 Tran zeatin (tZ) 0 ± 0 According to Table 3 , three PGRs are most represented in F. vesiculosus . The salicylic acid was detected in large quantity with up to 112.7 µg kg − 1 of d.s., followed by auxin (53.5 µg kg − 1 d.s.) and the ethylene precursor – ACC (1-amino-cyclopropane-1-carboxylic acid) (36.9 µg/kg d.s.). 3.2 Fucus vesiculosus- derived extracts 3.2.1 Biochemical composition Figure 2 presents the dry matter extraction yield (Figure 2(a)) and the biochemical composition of the extracts (Figure 2 (b-f)) obtained from F. vesiculosus after EAE and WE, according to the operational pH of each enzyme (one protease P2 at pH 7, two different glucosidases G1 and G3 at respectively pH 4 and pH 5). Figure 2 . (a) Extraction yield expressed as the mass of dry matter extracted per 100 g of dry F. vesiculosus and biochemical composition of Fucus -derived extracts expressed in percentage of dry extract (d.e.) – (b) total sugars; (c) reducing sugars; (d) uronic acids (% of dry mass of extract (d.e.); (e) proteins; (e); (f) Total Phenolic Content (mg eq phloroglucinol PE g − 1 d.e.) mean ± SD; n = 3. Results obtained for the dry matter extraction yield (Fig. 2 (a)) showed that pH influences the recovery of compounds from F. vesiculosus . Significantly higher dry matter extraction yields were observed with WE at pH 4 and pH 7, showing an increase of 18% (p = 0.001) and 14% (p = 0.003) respectively, compared to WE at pH 5. Notably, only G3 enzyme enhanced the dry matter extraction yield, presenting a statistically significant increase of 13% (p = 0.006) over WE at pH 5. Among the enzymes tested, G1 achieved the highest dry matter extraction yield at 60%, outperforming both enzyme G3 (p = 0.026) and P1 (p = 0.024). Regarding total sugars (Fig. 2 (b)), only the enzyme G3 allowed statistical increase (+ 34%) in the extraction of these compounds in comparison with the corresponding WE at pH 5. The extract obtained with G3 contained the highest amount of total sugars (38 ± 4% of d.e.) in comparison with the two other enzymes G1 and P2 with statistically significant differences between these groups (i.e., G1 and G3 and P2) with results multiplied by 1.8 ( p 0.0008) and by 2 ( p 0.0004) respectively). For both glycosidases (G1 and G3) significant higher amounts of reducing sugars were obtained (Fig. 2 (c)) equal to 21% and 10% d.e., respectively with significant differences in comparison with WE at pH 4 and WE and pH 5. Finally, even if extracts obtained with G1 had lower content of total sugars in comparison with G3, they contained twice more reducing sugars in comparison with G3 ( p 0.0002). The uronic acid content (Fig. 2 (d)) varied between 7% and 10% d.e., with no significant differences among the different extracts, except for those with P2 at pH 7. These latter extracts had the lowest uronic acid content at 6.8 ± 0.4% d.e., while the WE pH 5 extracts had the highest at 9.5 ± 0.8% d.e. Regarding proteins, the conventional water extracts contained 39 ± 3% d.e. of proteins at pH 5, and this increased to 53 ± 3% d.e. at pH 4 (Fig. 2 (e)). Statistically significant differences were observed between WE at pH 4 and pH 5 (p = 0.004), and between WE at pH 7 and pH 5 (p = 0.005). Extracts obtained with G1 and P2 had higher protein content compared to G3, with statistically significant differences. Figure 2 (f) shows the total phenolic content (TPC) of the different extracts. The results indicate that the highest TPC was found for the extracts obtained by EAE carried out with P2 enzyme at pH 7 (116 ± 7 mg PE g − 1 d.e.) with twice more TPC in comparison with its WE at pH 7 (54 ± 11 mg PE g − 1 d.e.). On the other hand, extract produced with G1 enzyme at pH 4 yielded a higher TPC (71 ± 11 mg PE g − 1 d.e.) than its WE at the same pH (58 ± 11 mg PE g − 1 d.e). Finally, G3 enzyme extract at pH 5 showed a TPC content of 57 ± 8 mg PE g − 1 d.e while the WE extract at pH 5 yielded a TPC content of 50 ± 9 mg PE g − 1 d.e. 3.2.2 Multi-element analysis Table 4 summarizes the multi-element composition of extracts obtained from Fucus vesiculosus by EAE and WE. Whatever the method and conditions of extraction, the most abundant elements are sodium, potassium and sulphur. It appears that conventional WE most often leads to similar or higher contents in macro- and microelements in aqueous extracts in comparison with extracts produced with EAE. Indeed, only few results show that enzymes allow for increasing the content of elements in the extract (d.e.), for example the content of nitrogen. The highest nitrogen content in the dry mass of the extract was obtained when P2 or G1 were used, leading to higher quantities that their respective WE at same pH. Table 4 Multi-element composition of extracts produced from Fucus vesiculosus by WE (pH 4 and pH 5) or EAE (enzymes G1, G3, P2) (mg kg − 1 d.e.; mean ± SD; n = 2) Element WE pH 4 WE pH 5 WE pH 7 G1 pH 4 G3 pH 5 P2 pH7 Macroelements Sodium 43 852 ± 614 42 913 ± 996 44 225 ± 388 47 397 ± 538 39 635 ± 692 45 035 ± 676 Potassium 40 623 ± 325 39 643 ± 919 38 603 ± 46 37 861 ± 644 36 081 ± 589 39 005 ± 702 Sulphur 14 128 ± 48 14 465 ± 248 13 676 ± 11 13 534 ± 257 12 476 ± 73 12 627 ± 278 Magnesium 7 988 ± 19 7 223 ± 76 6 940 ± 38 7 544 ± 106 6 653 ± 65 6 694 ± 74 Nitrogen 4 878 ± 470 5 659 ± 40 5 330 ± 320 5 844 ± 210 5 767 ± 744 6 188 ± 288 Calcium 7 097 ± 38 4 980 ± 42 4 962 ± 42 6 469 ± 45 4 962 ± 2 4 805 ± 30 Phosphorus 644 ± 8 741 ± 1 782 ± 16 699 ± 6 752 ± 2 753 ± 9 Microelements Boron 180 ± 1 190 ± 1 192 ± 5 168 ± 2 181 ± 1 181 ± 3 Iron 184 ± 1 139 ± 5 101 ± 2 179 ± 2 162 ± 4 96 ± 1 Manganese 91 ± 2 86 ± 1 83 ± 2 87 ± 1 83 ± 1 76 ± 1 Zinc 22 ± 0 23 ± 0 14 ± 0 20 ± 0 23 ± 1 14 ± 0 Copper 3 ± 0 3 ± 0 2 ± 0 2 ± 0 5 ± 0 2 ± 0 3.2.3 Plant Growth Regulators Table 5 presents the content of PGRs in extracts obtained through WE and EAE. As observed for F. vesiculosus powder (Table 3 ), the three main PGRs present in F. vesiculosus extracts were SA, IAA and ACC. Regarding the comparison of PGRs detected in raw F. vesiculosus powder and extracts, Table 5 reveals that new PGRs were identified in the extracts, such as cZR and tZR which are precursors of cZ and tZ respectively. Conversely, DJA is not anymore detected in the extracts. Table 5 Plant Growth Regulators composition of the extracts produced from Fucus vesiculosus (µg kg − 1 d.e.; mean ± SD; n = 3) Plant growth regulators WE pH 4 WE pH 5 WE pH 7 G1 pH 4 G3 pH 5 P2 pH7 Auxin Indole-3-acetic acid (IAA) 62 ± 6 b 88 ± 4 b 325 ± 34 a 58 ± 4 b 105 ± 12 b 289 ± 35 a Salicylic acid (SA) 213 ± 8 b 245 ± 17 b 289 ± 0 a 177 ± 3 c 245 ± 10 b 297 ± 5 a 1-amino-1-cyclopropane-carboxylic acid (ACC) 86 ± 35 bc 103 ± 4 bc 255 ± 29 a 57 ± 10 c 165 ± 45 ab 207 ± 22 a Abscisic acid (ABA) 3 ± 0 b 3 ± 0 b 5 ± 0 a 2 ± 0 b 3 ± 0 b 4 ± 0 a Cytokinins Isopentenyladenosin (iPR) 2 ± 1 c 1 ± 0 c 1 ± 0 c 12 ± 2 a 2 ± 0 c 4 ± 0 b Isopentenyladenine (iP) 1 ± 0 ab 1 ± 0 ab 1 ± 0 ab 1 ± 0 b 1 ± 0 ab 1 ± 0 a Cis zeatin (cZ) < 0.1 < 0.1 < 0.1 28 ± 2 < 0.1 < 0.1 Tran zeatin riboside (tZR) < 0.025 < 0.025 < 0.025 3 ± 1 < 0.025 < 0.025 Cis zeatin riboside (cZR) < 0.05 < 0.05 < 0.05 2 ± 0 < 0.05 2 ± 0 Tran zeatin (tZ) < 0.05 < 0.05 2 ± 0 < 0.05 < 0.05 < 0.05 3.2.4. Biological anti-oxidant activity The half maximal Inhibitory Concentration IC 50 values resulting from the DPPH test for the various F. vesiculosus powder and extracts are presented in Table 6 . The IC 50 values across different extracts ranged between 177.3 and 212.6 µg mL − 1 and are not all significantly different, making challenging the comparisons between DPPH scavenging activity of EAE and WE extracts. The highest total phenolic content was observed in the protease extract at pH 7 (116.4 ± 6.8 mg PE g − 1 d.e.). Despite this, it did not exhibit the greatest antioxidant activity. Surprisingly, it was the WE at pH 4 that showed the highest antioxidant activity, despite having a lower total phenolic content (57.7 ± 11 mg PE g − 1 d.e.) with an IC 50 value of 177.3 µg mL − 1 . Table 6 Dry weight rate and antioxidant activity (IC 50 ) of the Fucus vesiculosus powder and extracts Sample n Dry weight rate (% f.w.) Total phenolic content (mg PE g − 1 d.e or d.s) Antioxidant activity (IC 50 µg mL − 1 ) Mean ± SD * Mean ± SD * Mean ± SD ** F. vesiculosus powder 6 91.0 ± 0.1 d 39.8 ± 9.8 b 309.2 ± 33.9 c WE pH 4 extract 9 1.2 ± 0.0 a 57.7 ± 11.0 abc 177.3 ± 14.4 a WE pH 5 extract 15 1.1 ± 0.0 a 50.2 ± 9.3 ab 189.6 ± 15.5 a WE pH 7 extract 9 1.0 ± 0.0 a 54.2 ± 11.4 abc 212.6 ± 3.3 b P2 pH 7 extract 6 1.1 ± 0.1 a 116.4 ± 6.8 d 188.2 ± 15.5 a G1 pH 4 extract 9 1.4 ± 0.0 b 70.7 ± 10.6 cd 195.0 ± 12.7 ab G3 pH 5 extract 15 0.7 ± 0.3 c 56.5 ± 8.0 acd 186.2 ± 13.5 a * One-way analysis of means, not assuming equal variances, p < 0.05 ** One-way analysis of means, not assuming equal variances, p < 0.05. IC 50 of ascorbic acid (control): 15.1 ± 1.1 µg mL − 1 4. Discussion 4.1 Characterization of Fucus vesiculosus powder Comparing the data obtained with existing literature is challenging. Indeed, various factors, including those related to the harvesting, as well as others such as biomass pre-treatment (such as drying and grinding processes), selection of biochemical analysis techniques, contribute to significant variations. The main parameters responsible of variations in the biochemical composition of seaweed, including Fucus vesiculosus , are related to the harvest season, geographical location, environmental conditions like wave exposure and intensity, salinity, light availability, and reproductive phase or intra- and inter-specific competition (Paiva et al. 2018). Based on Fig. 1 , 33% of the non-detected compounds could be attributed to sulfated groups linked with fucose-rich polysaccharides, such as fucans and fucoidans (Rodríguez-Jasso et al. 2013 , Obluchinskaya et al. 2022 ). Fucus species are known to be rich in minerals, proteins, antioxidant phloroglucinol-based phenolic compounds, and cell wall polysaccharides, including alginates and sulphated polysaccharides rich in fucose (fucoidan) (Bikovens et al. 2017 ; Paiva et al. 2018; Obluchinskaya et al. 2022 ; Rousseau et al. 2023 ). The main constituents of the brown seaweed fibers are three distinct polysaccharides, namely sulfated polysaccharides rich in fucose (fucoidan), alginic acid (consists of β-D-mannuronic (M) and α-L-glucuronic (G) acid units) and laminaran (Catarino et al. 2018 ). The contents may vary between 3.4 and 25.7% d.w. for fucoidan, 8.4 and 58.8% for alginic acid and 0.6 and 7.0% for laminaran. Obluchinskaya et al. ( 2022 ) showed the amounts of monosaccharides, phlorotannins, flavonoids, and the mineral composition significantly varied in Fucus vesiculosus with reproductive phase and geographic locations in different Arctic region. Paiva et al. (2018) also discovered seasonal and geographical differences in the nutritional composition (protein, carbohydrate, lipid and fatty acids, dietary fiber) and functional activity (total phenolic and flavonoid content, antioxidant activity) of Fucus spiralis. Carbohydrates are the most abundant element of Fucus spp., involving in different roles including storage, defense responses, development, cell polarity or tissues integrity (Deniaud-Bouet et al. 2017). Carbohydrates can vary from 34–66% (d.w.) in Fucus vesiculosus (Catarino et al. 2018 ). The 22% of neutral sugars determined in F. vesiculosus in our study is quite similar to the result presented by Bikovens et al. ( 2017 ). Collected in March 2017 in Jurmala (Latvia), the total neutral sugars content of F. vesiculosus were analyzed using the same method and presented 24.6% d.w. Raw Fucus collected in Galicia (Spain) is rich in both soluble (10.52 ± 0.31%) and insoluble (48.63 ± 1.05%) fractions of dietary fibers (Diaz-Rubio et al. 2009). Fucus vesiculosus from Portugal is rich in carbohydrates (56.4 ± 0.4g/100 g seaweed d.w.) with fibers representing the main contribution (45.0 ± 0.1g/100 g d.w.). Fucus vesiculosus stood out by its high content of insoluble fiber compared to soluble fiber (Neto et al. 2018 ). Proteins are the second most important compound in this F. vesiculosus powder with 17% of d.s. which is in accordance with the literature. Indeed, according to Fleurence ( 1999 ), most of the brown seaweeds industrially exploited (Laminaria digitata, Ascophyllum nodosum, Fucus vesiculosus and Himanthalia elongata) have a protein content lower than 15% d.w. It shows little variations, from 11% (Catarino et al. 2018 ), to 13% d.w. (Lorenzo et al. 2017), with up to 15% d.w. (Neto et al. 2018 ). Within the Fucaceae family, phlorotannins are the main phenolic compounds found in brown algae, making them a valuable source of polyphenols (Obluchinskaya et al. 2022 ). In fact, phlorotannins have been identified as the only phenolic compounds in F. vesiculosus (Catarino et al. 2017 ). The abundance of phlorotannins in brown marine algae, as documented in the literature, ranges from 1–14% of dry algal biomass (Ar Gall et al. 2015 ; Catarino et al. 2019 ; Maheswaria and Babu 2022 ). In our study, the total phenolic content in the raw F. vesiculosus consisted of approximately 4% of dry weight (39.8 ± 9.8 mg eq PE g − 1 d.s) which is in accordance with the literature. The mineral content in seaweed is very variable (8–40% d.w.). It is primarily attributed to the properties of their cell surface polysaccharides, which facilitate the retention of inorganic marine substances. In the particular case of Fucus spp., the ash (mineral) content may vary from 19–36% d.w. (Catarino et al. 2018 ). Obluchinskaya et al. in 2022 confirmed that the ash content of macroalgae varies significantly, depending on the species, geographic location, and reproductive phase. The authors showed averages ash values of 18.5% and 28.5% d.w. depending on reproductive phases and locations. In our study, Fucus vesiculosus contains low amount of minerals with only 12% of ashes (Fig. 1 ). Fucus vesiculosus contains several important minerals for plants such as P, K, Ca, Mg. According to Catarino et al. ( 2018 ), sodium and potassium are the major elements described in this genus. According to Balina et al. ( 2016 ), F. vesiculosus from Jurmala, Latvia contains 21 500 mg kg − 1 of calcium, 11 000 mg kg − 1 of potassium, 9 300 mg kg − 1 of magnesium, 6 300 mg kg − 1 of sodium, 1 680 mg kg − 1 of manganese, 1 400 mg kg − 1 of phosphorus, 490 mg kg − 1 of iron, 89 mg kg − 1 of zinc and 12.7 mg kg − 1 of copper. The amounts of iron and magnesium obtained in this study are quite similar by contrast, phosphorus, zinc, copper and manganese are much lower (by 49–94% respectively). In addition, calcium is present in lower quantity in comparison with the results of literature (lower by 26%), while potassium and sodium are present in higher quantity (increase by 136% and 305% respectively). These differences could be explained by the effect related to local harvesting conditions (i.e., Spain for Balina et al. ( 2016 ) and France in this study) and seasonal variations (i.e., in January for Balina et al. ( 2016 ) and in July in this study) as reported and confirmed by Allen et al. ( 2015 ). In addition, content of elements in seaweeds depend on the concentrations of metal compounds in the surrounding water and they ability to bioaccumulate such compounds (Adams et al. 2011 ). Following the initial discovery of auxin in marine algae by Overbeek in 1940, research attention shifted towards identifying Plant Growth Regulators (PGRs) present in seaweeds. However, conclusive identification of auxin remained uncertain until the 1970s. During the past two decades, various PGRs (auxins, cytokinins, gibberellins, abscisic acid) were reported in different seaweed extracts (Stirk and Van Staden 2014 ). The first mention of the potential presence of PGR-type compounds in F. vesiculosus dates back to 1971. Biological tests carried out on Avena coleoptiles revealed the release of several indole-type compounds by the alkaline hydrolysis of various seaweeds, including Fucus vesiculosus (Buggeln 1971 ). Most recently, a study determined the content of PGRs in F. vesiculosus (Sharma et al. 2012). The acidic extracts obtained from F. vesiculosus collected in June contained up to 46.84 µg per kilogram of d.s which is in the same range with the results in this study. Regarding other brown seaweed species, the profiles and contents of PGRs have been explored in Sargassum horneri (Li et al. 2016 ). The content of salicylic acid was two times lower and the amount of IAA (Indole-3-acetic acid) six times lower compared with the present study. These authors have detected 3.9 µg kg − 1 for iP (Isopentenyladenine); 2.4 µg kg − 1 of iPR (Isopentenyladenosine) and 5.6 µg kg − 1 of ABA (Abscisic acid). Yalcin et al. ( 2019 ) observed that the content of phytohormones is very low for two brown seaweeds from 3.5 µg kg − 1 for Cladostephus spongiosum to 15.4 µg kg − 1 for Cystoseira barbata . Stirk et al. ( 2003 ) showed a content of IAA of 2.04 µg kg − 1 for Ecklonia maxima and 1.7 µg kg − 1 for Macrocystis pyrifera . However, in the present work, iP, iPR, cZ and ABA contents are quite similar to the values published by these authors. Stirk et al. ( 2003 ) have measured an amount of iP at 0.042 µg kg − 1 for Ecklonia maxima and 0.035 µg kg − 1 for Macrocystis pyrifera ; iPR at 0.006 µg kg − 1 and 0.003 µg kg − 1 respectively and cZR was under the limit of detection for both seaweeds. In general, PGRs from seaweeds are extracted with an organic solvent (i.e., methanol) with or without sonication. Subsequent purification by SPE or affinity chromatography is usually applied and the identification and quantification of PGRs are then performed by LC-MS or GC-MS (Benitez-Gracia et al. 2020; Yalcin et al. 2019 ). In the present study, the PGRs from F. vesiculosus were purified after extraction and concentrated by C18 or cation-exchange SPE depending on the nature of the molecule of interest. The quantification was performed by UHPLC-QTRAP-MS/MS in MRM (Multiple reaction monitoring) mode to achieve high specificity and sensitivity. The combination of an advanced chromatographic separation technique with tandem mass spectrometry allows the detection of PGRs down to ng/kg. While there are a few studies that have determined the levels of PGRs in seaweed, the final quantification is evidently affected by the method of harvesting and processing the seaweed samples after collection, including whether or not the raw material is dried. 4.2 Comparison of Fucus vesiculosu s Extracts Derived from Conventional Aqueous Extraction (WE) and Enzyme-Assisted Extraction (EAE) Regarding total sugars (Fig. 2 (b)), it was expected that extracts obtained with glycosidases would contain more sugars compared with protease due to the main activity dedicated to hydrolysis of osidic compounds such as oligo and polysaccharides. Results demonstrated the efficient enzymatic hydrolysis of polysaccharides contained in the seaweed thanks to G3 which allowed an increase in total sugars. As a result, the extract obtained using G3 at pH 5 was richer in sugars compared to the conventional water extraction method. Although EAE can significantly enhance the extraction of polysaccharides from brown algae by breaking down the cell wall, no studies were found in the literature specifically comparing our results obtained with the glucosidase enzyme. It is possible to assume that even if G1 enzyme breakdowns more sugars molecules, such as polysaccharides, resulting in more reducing ends (Fig. 2 (c)), the obtained polysaccharides are not sufficiently hydrolyzed to be quantified by the colorimetric method used here for the determination of total sugars. Seaweed-derived polysaccharides have the potential to act as plant biostimulants. Mzibra et al. ( 2021 ) showed that polysaccharide enriched extract obtained from Fucus spiralis significantly increased germination percentage, germination speed, reduced mean germination time of tomato seeds treated with this extract at a dose of 0.02 mg/mL. Moreover, the same extract applied to the soil and as a foliar spray enhanced tomato growth parameters such as leaves number, plant length, dry weight of shoot and root, and chlorophyll content. Polysaccharide enriched extract from F. spiralis applied to the soil improved plant growth parameters more successfully than applied as a foliar spray. Regarding proteins (Fig. 2 (e)), None of the tested enzymes allowed the recovery of more proteins in comparison with their respective WE, even for the protease P2. Agregan et al. (2017) have analyzed the protein content of F. vesiculosus water extracts obtained through UAE. They obtained results higher than those presented in this study with 62.1 ± 2.3% d.e. A significant difference between the TPC (Fig. 2 (f)) was found for the EAE extraction with P2 enzyme compared with WE at the different tested pH. Among these extractions, the EAE with P2 enzyme at pH 7 seems to be the most efficient for enriching total phenolic content in F. vesiculosus extracts. Our findings align with the existing literature data. In fact, several studies have demonstrated that EAE can enhance the extraction efficiency of polyphenols from brown algae (Hardouin et al. 2014 ; Teixeira-Guedes et al. 2023 ). Specifically, the disruption of the algal cell wall during EAE leads to the release of intracellular constituents, including proteins, which are prone to complex with polyphenols, causing aggregation and eventual precipitation (Siriwardhana et al. 2008 ). These polyphenols may interact with proteins through various mechanisms such as hydrogen bonding, π-bonding, hydrophobic interactions, as well as ionic and covalent linkages. Wang et al. (2020) investigated the application of proteases to enhance the yield of polyphenols and antioxidants from the red seaweed Palmaria palmata . They observed an increase in the extraction efficiency of phenolic compounds following protease treatment, attributing this improvement to the conversion of proteins into small peptides and free amino acids. Indeed, the protease degradation of proteins can liberate bound polyphenols, thereby improving access to polyphenols and facilitating their extraction (Wang et al. 2020). According to Table 4 , Enzyme G1 increases phosphorus contents in enzymatic extracts in comparison with WE at pH 4 (increase by 9% respectively). The enzyme G3 permitted to increase the extraction of iron (increase by 17%) and copper (by 45%) in comparison with WE at pH 5. The comparison of the three enzymes shows that G1 allowed for improved extraction of calcium, iron, magnesium, manganese, sodium and sulphur reaching the highest content in the dry extract in comparison with the extraction with G3 and P2. On the other hand, enzyme G3 permitted to obtain the highest extraction of boron, copper, phosphorus and zinc in comparison with enzymes G1 and P2. P2 permitted to obtain the highest content of potassium and nitrogen in the extract in comparison with G1 and G3 and had an equivalent amount of phosphorus in the extract as for the enzyme G3. Finally, considering the three main macroelements mandatory for plant nutrition and growth (nitrogen, phosphorus and potassium), extracts obtained with P2 are promising with the highest amount of nitrogen 6188 ± 288 mg kg − 1 d.e., which can be correlated to the use of a protease. Plants require specific essential minerals crucial for their structural integrity and metabolic processes. Inadequate supply of these elements can result in severe abnormalities in plant growth and development. Among the macroelements, nitrogen, phosphorus, and potassium are primary nutrients found in both mineral and organic fertilizers. Nitrogen deficiency adversely impacts plant productivity by reducing photosynthesis, leaf area, and the lifespan of green foliage. Phosphorus deficiency limits plant growth and crop yield, while potassium deficiency makes plants more susceptible to stress, leading to stomatal closure and inhibition of photosynthetic rates in various crop species (Wissuwa et al. 2003; Wang et al. 2013 ; Mu et al. 2021). The current study demonstrates that EAE extracts contain a range of elements, including nitrogen, phosphorus, and potassium. Therefore, application of these extracts on the crops could supply essential minerals for healthy growth (Choulot et al. 2022 ). Furthermore, these elements may play a role in activating plant defense mechanisms (Shukla et al. 2016). Literature data indicate that extract obtained from F. vesiculosus by UAE, rich in mineral components, may have a positive effect on plant growth, increasing the plant length, weight of plants, and chlorophyll content, in comparison with the control group (Krautforst et al. 2023 ). Michalak and Baśladyńska ( 2021 ) showed that the application of 5% F. vesiculosus extract, produced by hydrolysis with potassium hydroxide, on garden cress improved not only plant length and chlorophyll content, but also biofortified cultivated plants in Cu, Fe, Mn, Zn, Ca, S, P and K. The increases of some PGRs (Table 5 ) such as iPR and cZ in the dry extracts obtained through EAE with G1 pH4 could be explained by a more efficient and specific breakdown of seaweed cell wall polymers, thus improving the release of these compounds. This is implied by the increase in reducing sugars in G1 and G3 extracts in comparison with the WE (Fig. 2 (c)). In this way, the selective depolymerization of cell wall polysaccharides and proteins tends to increase the diffusion of PGRs into the soluble phase, thus leading to higher contents for some PGRs. Moreover, two additional PGRs were quantified in the extracts (WE and EAE) in comparison with the raw seaweed. This could be explained by the temperature of extraction, the pH or the longer duration of extraction allowing a better cell wall disruption. Regarding SA, the highest amount in the extract was obtained at pH 7 (WE and EAE) with significant differences observed with extracts obtained at pH 4 and pH 5. The same tendency was observed for IAA, ACC and ABA leading to the conclusion that an acidic pH has a negative effect on the recovery of these PGRs. About cytokinins, the enzymes G1 and P2 allowed statistically significant increases in the content of iPR in comparison with extract produced by WE under the same extraction conditions. The largest increase in the content of iPR was observed in the extract obtained using the enzyme G1, which contained seven times more iPR ( p 0.001) compared with WE pH 4. For cZ, tZ and their precursors, in most of the extracts the values were below the lower limits of quantification of the analytical method (< 0.1; 0.05; 0.025 µg/kg d.e.), except for the enzyme G1 which enabled the production of extracts with quantified amounts of cZ, cZR and Tz. If the enzyme efficiency is compared, it appears that the extract obtained with the protease is richer in PGRs than extracts obtained with glycosidases. Indeed, P2 allowed to increase the content of iP by 8%, SA by 21%, ACC by 26%, ABA by 36%, iPR by 132% and IAA by 175% in extracts in comparison with the G3 extract. Results obtained for the brown seaweed F. vesiculosus in this study were in the range to those obtained by Wally et al. ( 2012 ) for the commercial extracts produced from brown seaweeds collected in different countries, including Ascophyllum nodosum from France. The authors observed that the content of iP varied, ranging from 27.7 µg kg − 1 d.e. for South Pacific Durvillea to 1 µg/kg for Ascophyllum nodosum. It is not detected for the South African Ecklonia extract. cZ was detected at 1.7 µg kg − 1 in N. Pacific Macrocystis extract, while it was not detected in extracts from other countries, including A. nodosum extract. The concentration of tZ was 10 µg/kg for Chinese Sargassum extract and 1 µg kg − 1 d.e. for South African Ecklonia , with no detection in extracts from other countries, including Ascophyllum extract. iPR level ranged from 58.4 µg kg − 1 for Irish soluble seaweed extract powder to non-detected for extracts from various other countries, including Ascophyllum extract. As for ABA, it ranged from 10 µg kg − 1 for Chinese Sargassum extract to non-detected for extracts from various other countries, including French Ascophyllum extract. The content of IAA ranged from 615 µg kg − 1 for Norwegian extract to 6 µg kg − 1 for A. nodosum . Based on this study, the amount of IAA in A. nodosum extracts seems to be strongly influenced by the harvesting location (country/continent) (Wally et al. 2012 ). Another study analyzed the PGRs contents in commercial extracts produced from French Atlantic brown seaweed A. nodosum (Wally et al. 2012 ). The IAA and iP contents were 6 µg kg − 1 d.e. and 1 µg kg − 1 d.e., respectively, which are much lower than the values reported in the present study. The fact remains that A. nodosum has been recognized for many decades for its fertilizing and biostimulant properties. Thus, it appears that F. vesiculosus holds promise as a raw material to develop seaweed-based product for enhancing plant growth and development. This potential stems from the diverse properties of several growth regulators found within these extracts, including their ability to improve germination, root development, and flowering, thereby supporting crucial cellular processes such as iodine uptake and energy flow within plants (Wally et al. 2012 ; Benkova 2016). Moreover, these regulators create conducive environments for plant growth, metabolism, and insect pollination (Koo et al. 2020 ). Mattner et al. ( 2018 ) conducted a study where a commercial seaweed extract derived from Durvillaea potatorum and A. nodosum was applied to strawberries during both nursery and production stages. Their findings demonstrated an increase in root length and density of plants during the production stage with the use of seaweed extracts. Consequently, Mattner et al. ( 2018 ) suggested that seaweed extracts may enhance plant water and nutrient utilization, implying that components of seaweed other than minerals, such as PGRs, may be involved in this process. Specifically, IAA, a major PGRs present in these extracts, acts as a growth stimulant, promoting increased plant size (Benkova et al. 2016). Additionally, ABA functions as a growth inhibitor, inducing seed dormancy and influencing processes such as floral transition, lateral root formation, and seedling growth (Yang et al. 2022). Salicylic Acid, on the other hand, plays crucial roles in regulating plant disease resistance, thermogenesis, abiotic stress tolerance, DNA damage/repair, fruit yield, seed germination, and other physiological processes (Dempsey et al. 2017). 4.3 Evaluation of antioxidant activity with DPPH Our findings presented in Table 6 did not reveal a straightforward relationship between antioxidant activity and TPC in F. vesiculosus extracts. This result aligns with Sanchez-Bonnet et al. (2021) who described a significant negative correlation between TPC and DPPH radical scavenging activity in F. vesiculosus extracts (conventional and ultrasonic assisted extraction). They found that many of their extracts containing higher and lower content of polyphenols gave lower and higher antioxidant activity, respectively (Sanchez-Bonnet et al. 2021). Similarly, Hermund et al. ( 2018 ) also reported a negative correlation between TPC and RSA for F. vesiculosus aqueous ethanol extracts. They proposed that the lack of a positive correlation suggests that the extraction process was not selective, and other co-extracted compounds, such as polysaccharides, pigments, proteins, peptides, or vitamins, may influence the overall antioxidant activity synergistically or antagonistically. However, it is important to emphasize that F . vesiculosus powder showed a total phenolic content of 39.8 ± 9.8 mg PE g − 1 d.s. and exhibited the lowest antioxidant activity with an IC 50 of 309.2 µg mL − 1 compared to the different extraction techniques applied in this study. Since the total phenolic content found with P2 enzyme at pH 7 was significantly greater than the content in the seaweed powder, this suggests that EAE with P2 not only enhance the overall polyphenol content in the extracts, but also improve the antioxidant activity in F. vesiculosus by lowering their IC 50 value. 5. Conclusion This study aimed to develop and evaluate the effectiveness of two extraction methods — Enzyme Assisted Extraction (EAE) and Water Extraction (WE) — for the valorization of Fucus vesiculosus Linnaeus. The findings indicated that pH influenced the recovery of compounds from F. vesiculosus , with EAE showing higher efficiency at pH 4 compared to pH 5 and pH 7. Although EAE at pH 4 had a higher extraction yield than WE at the same pH, the difference was not significant. Similarly, WE was more efficient at pH 4 than at pH 5 and pH 7. Furthermore, this study demonstrated that EAE was significantly more effective than WE in producing enriched extracts, enhancing the release of bioactive compounds such as total sugars, a few plant growth regulators (PGRs), and polyphenols. Additionally, the polyphenol content and antioxidant activity of the raw algal powder and the various extracts were compared. The results highlighted that the extracts exhibited superior antioxidant activity, which can play a crucial role in agriculture by enhancing plant health through protection against oxidative stress, thereby supporting sustainable and eco-friendly agricultural practices by reducing reliance on chemical inputs. Declarations Acknowledgments This research was supported by the National Science Centre in Poland [grant 2019/33/B/NZ9/01844]; the French government under the programme “Investissements d'Avenir” - France 2030; Agro Innovation International – Timac Agro; and the ISblue project [grant ANR- 17-EURE-0015]. The authors thank Damien Souquet for the LC-MS analyses of PGRs. Ethical Approval Not applicable Funding This research was supported by the National Science Centre in Poland [grant 2019/33/B/NZ9/01844]; the French government under the programme “Investissements d'Avenir” - France 2030; Agro Innovation International – Timac Agro; and the ISblue project [grant ANR- 17-EURE-0015]. Availability of data and materials The data that support the findings of this study are available from the corresponding author, (Nathalie Bourgougnon). Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. References Adams, JMM, Ross AB, Anastasakis K, Hodgson E.M, Gallagher JA, Jones JM, Donnison IS (2011) Seasonal variation in the chemical composition of the bioenergy feedstock Laminaria digitata for thermochemical conversion. Bioresource Technol, Special Issue: Biofuels - II: Algal Biofuels and Microbial Fuel Cells 102:226–234. https://doi.org/10.1016/j.biortech.2010.06.152 Agregán R, Munekata PE, Domínguez R, Carballo J, Franco D, Lorenzo JM (2017) Proximate composition, phenolic content and in vitro antioxidant activity of aqueous extracts of the seaweeds Ascophyllum nodosum, Bifurcaria bifurcata and Fucus vesiculosus. Effect of addition of the extracts on the oxidative stability of canola oil under accelerated storage conditions. Food Res Int 99:986–994. https://doi.org/10.1016/j.foodres.2016.11.009 Allen E, Wall DM, Herrmann C, Xia A, Murphy JD (2015) What is the gross energy yield of third generation gaseous biofuel sourced from seaweed? Energy 81:352–360. https://doi.org/10.1016/j.energy.2014.12.048 Arzel P (1987) Les Goémoniers. Le Chasse-Marée, Douarnenez. ISBN 2903708053 Balina K, Romagnoli F, Blumberga D (2016) Chemical Composition and Potential Use of Fucus vesiculosus from Gulf of Riga. Energy Proced 95:43–49. https://doi.org/10.1016/j.egypro.2016.09.010 Benková E (2016) Plant hormones in interactions with the environment. Plant Mol Biol 91:597. https://doi.org/10.1007/s11103-016-0501-8 Benitez García I, Dueñas Ledezma AK, Martínez-Montaño E., Salazar Leyva JA, Carrera E, Osuna Ruiz I (2020) Identification and Quantification of Plant Growth Regulators and Antioxidant Compounds in Aqueous Extracts of Padina durvillaei and Ulva lactuca. Agronomy, 10: 866. https://doi.org/10.3390/agronomy10060866 Besednova NN, Andryukov BG, Zaporozhets TS, Kryzhanovsky SP, Kuznetsova TA, Fedyanina LN, Makarenkova ID, Zvyagintseva TN (2020) Algae polyphenolic compounds and modern antibacterial strategies: Current achievements and immediate prospects. Biomedicines 8:342. https://doi.org/10.3390/biomedicines8090342 Blumenkrantz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem. 54(2):484–9. https://doi.org/10.1016/0003-2697(73)90377-1 Bikovens O, Ponomarenko J, Janceva S, Lauberts M, Vevere L, Telysheva G (2017) Development of the Approaches for Complex Utilization of brown algae ( Fucus vesiculosus ) Biomass for the Obtaining of Value-Added Products. Proceedings of the 8th International Scientific Conference Rural Development. https://doi.org/10.15544/RD.2017.091 Buggeln RG, Craigie JS (1971) Evaluation of evidence for the presence of indole-3-acetic acid in marine algae. Planta 97:173–178. https://doi.org/10.1007/BF00386764 Bunker F, Brodie JA, Maggs CA, Bunker AR (2017) Seaweeds of Britain and Ireland: Second Edition. Wild nature press. ISBN 9780995567337 Catarino MD, Silva AMS, Cardoso SM (2017) Review Fucaceae: A Source of Bioactive Phlorotannins. Int J Mol Sci 18:1327. https://doi.org/10.3390/ijms18061327 Catarino, MD, Silva, AMS, Cardoso SM (2018) Phycochemical constituents and biological activities of Fucus spp. Mar Drugs 16:249. https://doi.org/10.3390/md16080249 Catarino MD, Silva AMS, Mateus N, Cardoso SM (2019) Optimization of phlorotannins extraction from Fucus vesiculosus and evaluation of their potential to prevent metabolic disorders. Mar Drugs 17(3):162. https://doi.org/10.3390/md17030162 Chambers JM, Hastie TJ (1992) Statistical Models in S, Wadsworth & Brooks/Cole Choulot M, Le Guillard C, Bourgougnon N, Michalak I (2022) Seaweeds-based fertilizing products in: “Algae and Aquatic Macrophytes in Cities”. Vimal Chandra Pandey, ISBN: 978-0-12-824270-4. Choulot M, Michalak I, Jing L, Szymczycha-Madeja A, Wełna M, Bourgougnon N, Le Guillard C (2023) The Enzyme-Assisted Extraction of compounds of interest in agriculture: Case study of the red seaweed Solieria chordalis (C. Agardh) J. Agardh, Algal Research 75:103239. https://doi.org/10.1016/j.algal.2023.103239 Dempsey DA, Klessig DF (2017) How does the multifaceted plant hormone salicylic acid combat disease in plants and are similar mechanisms utilized in humans? BMC Biol 15:23. https://doi.org/10.1186/s12915-017-0364-8 Deniaud-Bouët E, Hardouin K, Potin P, Kloareg B, Hervé, C (2017) A review about brown algal cell walls and fucose-containing sulfated polysaccharides: Cell wall context, biomedical properties and key research challenges. Carbohyd Polym 175:395–408. https://doi.org/10.1016/j.carbpol.2017.07.082 Díaz-Rubio ME, Pérez-Jiménez J, Saura-Calixto F (2009) Dietary fiber and antioxidant capacity in Fucus vesiculosus products. Int J Food Sci. Nutr 60:23–34. https://doi.org/10.1080/09637480802189643 DuBois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric Method for Determination of Sugars and Related Substances. Anal Chem 28:350–356. https://doi.org/10.1021/ac60111a017 Filisetti-Cozzi TMCC, Carpita NC (1991) Measurement of uronic acids without interference from neutral sugars. Anal Biochem 197: 157–162. https://doi.org/10.1016/0003-2697(91)90372-Z Fleurence J (1999) Seaweed proteins: biochemical, nutritional aspects and potential uses. Trends Food Sci Tech 10:25–28. https://doi.org/10.1016/S0924-2244(99)00015-1 Fox J, Bouchet-Valat M, Marquez MM, et al. (2024) Rcmdr: R Commander. https://cran.r-project.org/web/packages/Rcmdr/index.html ; accessed 8 February 2024 Gall EA, Lelchat F, Hupel M, Jégou C, Stiger-Pouvreau V (2015) Extraction and Purification of Phlorotannins from Brown Algae. In: Stengel, D., Connan, S. (eds) Natural Products From Marine Algae. Methods in Molecular Biology, vol 1308. Humana Press, New York. https://doi.org/10.1007/978-1-4939-2684-8_7 Haddad C, Arkoun M, Jamois F, Schwarzenberg A, Yvin J-C, Etienne P and Laîné P (2018) Silicon promotes growth of Brassica napus L. and delays leaf senescence induced by nitrogen starvation. Front Plant Sci. 9:516. https://doi.org/10.3389/fpls.2018.00516 Hagerman AE, Butler LG (1989) Choosing appropriate methods and standards for assaying tannin. J Chem Ecol 15:1795–1810. https://doi.org/10.1007/BF01012267 Hardouin K, Bedoux G, Burlot AS, Nyvall-Collen P, Bourgougnon N (2014) Enzymatic recovery of metabolites from seaweeds: potential applications. Adv Bot Res 71:279–320. https://doi.org/10.1016/B978-0-12-408062-1.00010-X Hardouin K, Burlot AS, Umami A, Tanniou A, Stiger-Pouvreau V, Widowati I, Bedoux G, Bourgougnon N (2014) Biochemical and antiviral activities of enzymatic hydrolysates from different invasive French seaweeds. J Appl Phycol 26:1029–1042. https://doi.org/10.1007/s10811-013-0201-6 Hermund DB, Plaza M, Turner C, Jónsdóttir R, Kristinsson HG, Jacobsen C, Jacobsen C, Fog Nielsen K (2018) Structure dependent antioxidant capacity of phlorotannins from Icelandic Fucus vesiculosus by UHPLC-DAD-ECD-QTOFMS. Food Chem 240:904–909. https://doi.org/10.1016/j.foodchem.2017.08.032 Hollander M, Wolfe DA, Chicken E (1973). Nonparametric Statistical Methods. Wiley, New York Jacob C, Frangoudes K, Mesnildrey L, Lesueur M, Spinec F, Folliard G, Pien S, Rebours C, Meland M, Gallastegi I, Inunziaga Urizarbarrena A, Marques A, Atack T, Dunningham J, Maguire J, Abreu H, Walsh M (2012) Etats des lieux de la filière des macro-algues en Europe. projet NETALGAE - Interreg IVB. World Bank (2023) Global Seaweed: New and Emerging Markets Report, 2023. © Washington, DC:World Bank. http://hdl.handle.net/10986/40187 Kadam SU, Tiwari BK, O’Donnell CP (2013) Application of novel extraction technologies for bioactives from marine algae. J. Agric. Food Chem. 61:4667–4675. https://doi.org/10.1021/jf400819p Knight M, Parke M (1950) A biological study of Fucus vesiculosus L. and Fucus serratus L . J Mar Biol Assoc UK 29:439–514. https://doi.org/10.1017/S0025315400055454 Koo YM, Heo AY, Choi HW (2020) Salicylic Acid as a Safe Plant Protector and Growth Regulator. Plant Pathol J 36(1):1–10. https://doi.org/10.5423/PPJ.RW.12.2019.0295 Krautforst K, Szymczycha-Madeja A, Welma M, Michalak I (2023) Brown seaweed: Fucus vesiculosus as a feedstock for agriculture and environment protection. Sci Rep 13, 100065 https://doi.org/10.1038/s41598-023-36881-z Laekeman, G (2014) Assessment Report on Fucus vesiculosus L., Thallus https://www.ema.europa.eu/en/documents/herbal-report/final-assessment-report-fucus-vesiculosus-l-thallus_en.pdf Le Guillard C, Bergé, JP, Donnay-Moreno C, Cornet J, Ragon JY, Fleurence J, Dumay J (2023) Optimization of R-Phycoerythrin Extraction by Ultrasound-Assisted Enzymatic Hydrolysis: A Comprehensive Study on the Wet Seaweed Grateloupia turuturu . Mar Drugs 21:213. https://doi.org/10.3390/md21040213 Le Lann K, Jégou C, Stiger-Pouvreau V (2008) Effect of different conditioning treatments on total phenolic content and antioxidant activities in two Sargassacean species: Comparison of the frondose Sargassum muticum (Yendo) Fensholt and the cylindrical Bifurcaria bifurcata R. Ross. Phycol Res 56: 238–245. https://doi.org/10.1111/j.1440-1835.2008.00505.x Li J, Chen K, Li S, Liu T, Wang F, Xia Y, Lu J, Zhou Y, Guo C (2016) Pretreatment with Fucoidan from Fucus vesiculosus Protected against ConA-Induced Acute Liver Injury by Inhibiting Both Intrinsic and Extrinsic Apoptosis. PLOS One 11(4):e0152570. https://doi.org/10.1371/journal.pone.0152570 Maheswaria V, Babu PA S (2022) Phlorotannin and its derivatives, a potential antiviral molecule from brown seaweeds, an overview. Russ J Mar Biol + 48 (5):309–324. https://doi.org/10.1134/S1063074022050169 Mahmoud E, Abd El-Kader N (2015) Heavy metal immobilization in contaminated soils using phosphogypsum and rice straw compost. Land Degrad Dev 26:819–824. https://doi.org/10.1002/ldr.2288 Mattner SW, Milinkovic M, Arioli T (2018) Increased growth response of strawberry roots to a commercial extract from Durvillaea potatorum and Ascophyllum nodosum . J Appl Phycol 30:2943–2951. https://doi.org/10.1007/s10811-017-1387-9 Mesnildrey L, Jacob Céline, Frangoudes K, Reunavot M, Lesueur M (2012) Seaweed industry in France. Report Interreg program NETALGAE. 2012, 42 p. ⟨hal-00840572⟩ Michalak I, Chojnacka K (2014) Algal extracts: Technology and advances. Eng Life Sci 14: 581–591. https://doi.org/10.1002/elsc.201400139 Michalak I, Baśladyńska S (2021) Effect of Fucus extract and biomass enriched with Cu (II) and Zn (II) ions on the growth of garden cress (Lepidium sativum) under laboratory conditions. Ital J Agron 16: 1799. https://doi.org/10.4081/ija.2021.1799 Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31(3):426–428. https://doi.org/10.1021/ac60147a030 Mu X, Chen Y (2021) The physiological response of photosynthesis to nitrogen deficiency. Plant Physiol Bioch 158:76–82. https://doi.org/10.1016/j.plaphy.2020.11.019 Mzibra, A., Aasfar, A., Benhima, R. et al. Biostimulants derived from Moroccan seaweeds: seed germination metabolomics and growth promotion of tomato plant. J Plant Growth Regul 40, 353–370 (2021). https://doi.org/10.1007/s00344-020-10104-5 Neiva J, Serrão EA, Assis J, Pearson GA, Coyer JA, Olsen JL, Hoarau G, Valero M (2016) Climate oscillations, range shifts and phylogeographic patterns of North Atlantic Fucaceae. In: Hu Z-M, Fraser C (eds) Seaweed Phylogeography. Springer Netherlands, Dordrecht, pp279–308 Neto RT, Marçal C, Queirós AS, Abreu H, Silva AMS, Cardoso SM (2018) Screening of Ulva rigida , Gracilaria sp., Fucus vesiculosus and Saccharina latissima as Functional Ingredients. Int J Mol Sci 19:2987. https://doi.org/10.3390/ijms19102987 Obluchinskaya ED (2008) Comparative chemical composition of the Barents Sea brown algae. Appl Biochem Microbiol 44:305–309. https://doi.org/10.1134/S0003683808030149 Obluchinskaya ED, Pozharitskaya ON, Zakharov DV, Flisyuk EV, Terninko II, Generalova YE, Smekhova IE, Shikov AN (2022) The Biochemical Composition and Antioxidant Properties of Fucus vesiculosus from the Arctic Region. Mar Drugs 20:193. https://doi.org/10.3390/md20030193 Ososki AL, Lohr P, Reiff M, Balick MJ, Kronenberg F, Fugh-Berman A, O’Connor B (2002) Ethnobotanical literature survey of medicinal plants in the Dominican Republic used for women’s health conditions. J Ethnopharmacol 79:285–298. https://doi.org/10.1016/S0378-8741(01)00376-2 Overbeek, JV (1940) Auxin in Marine Algae. Plant Physiol 15:291–299. https://doi.org/10.1104/pp.15.2.291 Paiva L., Lima E., Neto A.I., Baptista J. Seasonal variability of the biochemical composition and antioxidant properties of Fucus spiralis at two Azorean islands. Mar. Drugs 2018, 16, 248; https://doi.org/10.3390/md16080248 Parys S, Kehraus S, Krick A, Glombitza KW, Carmeli S, Klimo K, König GM (2010) In vitro chemopreventive potential of fucophlorethols from the brown alga Fucus vesiculosus L. by anti-oxidant activity and inhibition of selected cytochrome P450 enzymes. Phytochemistry 71:221–229. https://doi.org/10.1016/j.phytochem.2009.10.020 Pierre G (2010). Biochemical characterization of algal exopolymers from Marennes-Oléron and study of the physico-chemical surface properties of microorganisms involved in their adhesion (phdthesis). Université de La Rochelle. Réthoré E, Jing L, Ali N, Yvin JC, Pluchon S, Hosseini SA (2021) K Deprivation modulates the primary metabolites and increases putrescine concentration in Brassica napus . Front Plant Sci 12:681895. https://doi.org/10.3389/fpls.2021.681895 Rodríguez-Jasso RM, Mussatto SI, Pastrana L, Aguila CN, Teixeira JA (2013) Extraction of sulfated polysaccharides by autohydrolysis of brown seaweed Fucus vesiculosus . J Appl Phycol 25:31–39. https://doi.org/10.1007/s10811-012-9834-0 Romm A, Hardy ML, Mills S, Bove M, Stansbury JE, Romm A (2010) Chap. 6 Endocrine Disorders and Adrenal Support. In: Botanical Medicine for Women’s Health; Churchill Livingstone: London, UK, pp 186–210. https://doi.org/10.1016/B978-0-443-07277-2.X0001-3 Rousseau C, Demoulinger G, Rousvoal S, Champeval D, Dolly M, Michel G, Thomas F, Markov G.V, Potin P, Dittami S.M, Prado S, Leblanc C, Le Guillard C (2023) A review on the chemical ecology of the Fucaceae holobionts: from fundamental knowledge to applications. CR Chim 26:1–25. https://doi.org/10.5802/crchim.271 Saebmehr H, Rafiee F, Givianrad M H, Mostafavi G (2022) Determination and quantification of two regulatory phytohormones extracted from Sargassum muticum and Gracilaria corticate seaweeds in the south of Iran. Nova Biologica Reperta 9(2):115–123. https://doi.org/10.29252/nbr.9.2.115 Sanchez-Bonet D, García-Oms S, Belda-Antolí M, Padrón-Sanz C, Lloris-Carsi JM, Cejalvo-Lapeña D (2021) RP-HPLC-DAD determination of the differences in the polyphenol content of Fucus vesiculosus extracts with similar antioxidant activity. J Chromatogr B 1184:122978. https://doi.org/10.1016/j.jchromb.2021.122978 Sharma HSS, Lyons G, McRoberts C, McCall D, Carmichael E, Andrews F, Swan R (2012a) Biostimulant activity of brown seaweed species from Strangford Lough: compositional analyses of polysaccharides and bioassay of extracts using mung bean (Vigno mungo L.) and pak choi (Brassica rapa chinensis L.). J Appl Phycol 24:1081–1091. https://doi.org/10.1007/s10811-011-9737-5 Sharma HSS, Fleming C, Selby C, Rao JR, Martin T (2014) Plant biostimulants: a review on the processing of macroalgae and use of extracts for crop management to reduce abiotic and biotic stresses. J Appl Phycol 26:465–490. https://doi.org/10.1007/s10811-013-0101-9 Singleton VL, Orthofer R, Lamuela-Raventos RM (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu Reagent. Method Enzymol 299: 152–178. https://doi.org/10.1016/S0076-6879(99)99017-1 Siriwardhana N, Kim KN, Lee KW, Kim SH, Ha JH, Song CB, Lee JB, Jeon YJ (2008) Optimisation of hydrophilic antioxidant extraction from Hizikia fusiformis by integrating treatments of enzymes, heat and pH control. Int J Food Sci Tech 43(4):587–596. https://doi.org/10.1111/j.1365-2621.2006.01485.x Skibola CF (2004) The effect of Fucus vesiculosus , an edible brown seaweed, upon menstrual cycle length and hormonal status in three pre-menopausal women: A case report. BMC Complement. Altern Med 4:10. https://doi.org/10.1186/1472-6882-4-10 Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem, 150(1):76–85. https://doi.org/10.1016/0003-2697(85)90442-7 Spain O, Hardouin K, Bourgougnon N, Michalak I (2024) Enzyme Assisted Extraction of red seaweed Solieria chordalis ) J. Agardh 184 - the starting point for the production of biostimulants of plant growth and biosorbents of metal ions. Biomass Conversion and Biorefinery Journal 14:1621–1635. https://doi.org/10.1007/s13399-022-02456-7 Shukla PS, Borza T, Critchley AT, Prithiviraj B (2021) Seaweed-Based Compounds and Products for Sustainable Protection against Plant Pathogens. Mar Drugs 19(2):59. https://doi.org/10.3390/md19020059 Stirk W, Novak O, Strnad M, Staden J (2003) Cytokinins in macroalgae. Plant Growth Regul 41:13–24. https://doi.org/10.1023/A:1027376507197 Stirk WA, Van Staden J (2014) Chapter Five: Plant Growth Regulators in Seaweeds: Occurrence, Regulation and Functions. In: Bourgougnon, N. (Ed.), Advances in Botanical Research, Sea Plants. Academic Press, pp 125–159. https://doi.org/10.1016/B978-0-12-408062-1.00005-6 Teixeira-Guedes C, Gomes-Dias JS, Cunha SA, Pintado ME, Pereira RN, Teixeira JA, Rocha CMR (2023) Enzymatic approach for the extraction of bioactive fractions from red, green and brown seaweeds. Food Bioprod Process 138:25–39. https://doi.org/10.1016/j.fbp.2022.12.005 Wally OSD, Critchley AT, Hiltz D, Craigie JS, Han X, Zaharia LI, Abrams SR, Prithiviraj B (2012) Regulation of Phytohormone Biosynthesis and Accumulation in Arabidopsis Following Treatment with Commercial Extract from the Marine Macroalga Ascophyllum nodosum. J Plant Growth Regul 32:324–339. https://doi.org/10.1007/s00344-012-9301-9 Wang M, Zheng Q, Shen Q, Guo S (2013) The Critical Role of Potassium in Plant Stress Response. Int J Mol Sci 14(4): 7370–7390. https://doi.org/10.3390/ijms14047370 Wang T, Jónsdóttir R, Kristinsson HG, Hreggvidsson GO, Jónsson JÓ, Thorkelsson G, Ólafsdóttir G (2010) Enzyme-enhanced extraction of antioxidant ingredients from red algae Palmaria palmata. LWT - Food Sci Technol 43:1387–1393. https://doi.org/10.1016/j.lwt.2010.05.010 Waterhouse AL (2002) Determination of Total Phenolics. Current Protocols in Food Analytical Chemistry, I1.1.1-I 1.1.8 . https://doi.org/10.1002/0471142913.fai0101s06 Wijesinghe WAJP, Jeon YJ (2012) Enzyme-assistant extraction (EAE) of bioactive components: A useful approach for recovery of industrially important metabolites from seaweeds: A review. Fitoterapia 83:6–12. https://doi.org/10.1016/j.fitote.2011.10.016 Wissuwa M (2003) How Do Plants Achieve Tolerance to Phosphorus Deficiency? Small Causes with Big Effects. Plant Physiol 133(4):1947–1958. https://doi.org/10.1104/pp.103.029306 Yalcin S, Okudan E, Karakaş Ö, Önem A, Sözgen Başkan K (2019) Identification and quantification of some phytohormones in seaweeds using UPLC-MS/MS. J Liq Chromatogr R T 42:1–10. https://doi.org/10.1080/10826076.2019.1625374 Additional Declarations No competing interests reported. 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Technologies","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Izabela","middleName":"","lastName":"Michalak","suffix":""},{"id":373978152,"identity":"813fb869-30d1-42a5-b192-b68e3b20153e","order_by":8,"name":"Nathalie Bourgougnon","email":"data:image/png;base64,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","orcid":"","institution":"Université Bretagne Sud, EMR CNRS, LBCM","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Nathalie","middleName":"","lastName":"Bourgougnon","suffix":""}],"badges":[],"createdAt":"2024-10-25 09:23:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5331195/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5331195/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":68452048,"identity":"d029e033-d76c-409c-b2b0-56108322f9f7","added_by":"auto","created_at":"2024-11-07 12:02:53","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":92161,"visible":true,"origin":"","legend":"\u003cp\u003eProximate composition of \u003cem\u003eFucus vesiculosus\u003c/em\u003eraw material (percentage of d.s.)\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5331195/v1/c8faa1fc2bbef7b382dd1f78.jpg"},{"id":68453134,"identity":"fc3200a9-1204-4f99-b5ff-c2fbec63f4b3","added_by":"auto","created_at":"2024-11-07 12:10:53","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":505240,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Extraction yield expressed as the mass of dry matter extracted per 100\u0026nbsp;g of dry \u003cem\u003eF.\u003c/em\u003e \u003cem\u003evesiculosus\u003c/em\u003e and biochemical composition of \u003cem\u003eFucus\u003c/em\u003e-derived extracts expressed in percentage of dry extract (d.e.) – (b) total sugars; (c) reducing sugars; (d) uronic acids (% of dry mass of extract (d.e.); (e) proteins; (e); (f) Total Phenolic Content (mg eq phloroglucinol PE g\u003csup\u003e-1\u003c/sup\u003e d.e.) mean±SD; n=3.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5331195/v1/8f0ba26d3b580d553c59b518.jpg"},{"id":68453420,"identity":"12e917fe-d450-4cb2-a470-55e15149033c","added_by":"auto","created_at":"2024-11-07 12:18:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1903211,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5331195/v1/da2c55a7-0810-41de-ba78-84bd33197520.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Application of Enzyme-Assisted Extraction on the brown seaweed Fucus vesiculosus Linnaeus (Ochrophyta, Fucaceae) to produce extracts for a sustainable agriculture.","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eOver the last few years, the agricultural sector is facing numerous challenges due to a burgeoning global population, climate warming, severe weather events, the over use of mineral fertilizers and in the same time the necessity to produce food in more efficient and sustainable ways. This context led again to a broader interest in natural alternatives to preserve environment and produce quality food. One of the most popular, intensively developing groups of biostimulants are seaweed extracts. Biostimulants are the most promising short-term markets for seaweed, projected to reach USD 4.4\u0026nbsp;billion by 2030. According to Global Seaweed New and Emerging Markets Report (2023), seaweed-based biostimulants can expect to see significant growth over the next few years as additional investment goes into research and development for improving efficacy, and as more and more seaweed processors are exploiting seaweed biomass as a whole in a biorefinery approach to create multiple added-value products.\u003c/p\u003e \u003cp\u003eFor centuries, coastal populations in various regions across the globe have harvested a diverse range of seaweeds, source of livelihood and survival (Jacob et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). These seaweeds were first used for domestic purposes, such as human health, consumption, or glass production (Arzel 1986). Seaweeds have also been employed as fertilizers, soil enhancers, and for soil pH adjustment. In regions such as Brittany and along the French Atlantic coast, seaweed was traditionally collected by coastal farmers using large rakes following storms and directly applied to fields. This traditional practice persists on the island of Batz in northern Brittany and the Rhuys peninsula in southern Brittany in France (Choulot et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The application of seaweed-based products in agriculture has been gaining momentum in crop production systems over the past 30 years. Today, seaweed used for biostimulants is mainly brown wild-harvested especially \u003cem\u003eDurvillaea potatorum\u003c/em\u003e and \u003cem\u003eEcklonia maxima\u003c/em\u003e (Laminariales) or \u003cem\u003eAscophyllum nodosum\u003c/em\u003e (Fucales) (Rousseau et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Various liquid products are present on the market such as Seasol with \u003cem\u003eDurvillaea potatorum\u003c/em\u003e, Kelpak with \u003cem\u003eEcklonia maxima\u003c/em\u003e, or Fertileader, AlgaGreen and Maxicrop with \u003cem\u003eAscophyllum nodosum\u003c/em\u003e (Choulot et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Rich in wide variety of compounds \u0026ndash; macronutrients and micronutrients, alginates and oligosaccharides, Plant Growth Regulators (PGRs), vitamins, and osmolytes \u0026ndash; seaweeds extracts are used as agricultural input products in different forms, such as biofertilizers, liming materials, soil improvers, plant biostimulants, and fertilizing product blends (e.g., Bikovens et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Michalak and Baśladyńska, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Mzibra et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Choulot et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Krautforst et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Spain et al. \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong the harvested algae in France, \u003cem\u003eFucus vesiculosus\u003c/em\u003e Linnaeus, commonly known as bladder wrack, is most commonly found on sheltered rocky shores subject to some degree of disturbance, such as scour (Bunker et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). In Brittany, \u003cem\u003eF. vesiculosus\u003c/em\u003e is particularly abundant, forming extensive intertidal zones characteristic of the mid and low-mid intertidal zones. This species can grow to sizes ranging from tens centimeters to one meter in length, with a lifespan typically between 2 and 5 years (Knight and Parke \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e1950\u003c/span\u003e). The biomass of \u003cem\u003eFucus\u003c/em\u003e can now reach several hundred to several thousand fresh tons per year (Mesnildrey et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; CSAVM 2023, personal communication), continuing to play a significant role in local economies for various industries such as cosmetics and agriculture. In agriculture, due to its unique chemical composition, Fucus sp. biomass is being tested for the use in agriculture as a soil additive, positively influencing plant growth, for example: oat (Bikovens et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), garden cress (Michalak and Baśladyńska, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) or sorghum (Krautforst et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFor the production of seaweed based biostimulants, alkaline extraction using potassium hydroxide is currently the most common method, constituting approximately 80% of all biostimulant production (Choulot et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Global Seaweed New and Emerging Markets Report 2023). However, some bioactive compounds such as PGRs, vitamins, and antioxidants are susceptible to degradation under harsh extraction conditions (extreme pH or/and high temperatures) (Sharma et al. \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The complexity of the cell wall, with strong interactions between fibrillar, matrix polysaccharides, polyphenols, and proteins, hampers the release of compounds during extraction. Numerous physicochemical and biological methods have been developed to recover active compounds from algal biomass (Kadam et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Michalak and Chojnacka, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). In the context of climate change and socio-environmental concerns, the blue circular economy, selecting an appropriate extraction method must balance metabolites selectivity and recovery, cost-effectiveness, and eco-friendly processes based on green and sustainable chemistry and engineering.\u003c/p\u003e \u003cp\u003eCurrently, Enzyme-Assisted Extraction (EAE), Microwaves Assisted Extraction (MAE) or Ultrasound-Assisted Extraction (UAE) are among the techniques most mentioned in the literature for natural biomass processing, used alone or combined (Wijesinghe and Jeon \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Michalak and Chojnacka, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Choulot et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Spain et al. 2022; Le Guillard et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Krautforst et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Choulot et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The processes are described as feasible and efficient alternatives to traditional extraction methods and might be transposed on a large scale for seaweed processing in some industrial sectors.\u003c/p\u003e \u003cp\u003eDue to the highly site-specific activity of the hydrolytic enzymes, enzymatic hydrolysis process has been an excellent, environmentally-friendly method to produce some active compounds. EAE effectively extracts valuable components from seaweed without the need for denaturing conditions such as solvents or high temperatures. The effectiveness of EAE in seaweed liquefaction has been demonstrated through the use of two types of enzymes: proteases and carbohydrases. Research has consistently shown that the use of enzymes can significantly enhance the value of seaweed extracts or compounds (Hardouin et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Michalak and Chojnacka, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Spain et al. 2022; Choulot et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). EAE is currently used in various industrial applications, including energy (methanation), agriculture, feed, and food (Michalak and Chojnacka, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Wijesinghe and Jeon, \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Terme et al., 2020; Choulot et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Several works explore EAE for the production of molecules of interest from seaweeds (Hardouin et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Obluchinskaya et al. 2021; Getachew et al. 2022; Spain et al. 2022; Pliego-Cortes et al. 2023) but no work reports the use of this technique for the production of biostimulants with \u003cem\u003eFucus vesiculosus\u003c/em\u003e biomass.\u003c/p\u003e \u003cp\u003eThis study explores the potential benefits of employing Enzyme-Assisted Extraction on \u003cem\u003eFucus vesiculosus\u003c/em\u003e Linnaeus (\u003cem\u003eOchrophyta\u003c/em\u003e, \u003cem\u003eFucaceae\u003c/em\u003e) by analyzing specific compounds pertinent to agriculture, including Plant Growth Regulators, polyphenols and macro/micronutrients.\u003c/p\u003e"},{"header":"2. Material and methods","content":"\u003cp\u003e \u003cem\u003eFucus vesiculosus\u003c/em\u003e Linnaeus powder (\u0026lt;\u0026thinsp;904 \u0026micro;m) was purchased from Groupe Roullier (Saint-Malo, France). Seaweeds were collected in Brittany (France) in July 2019 and dried in an industrial drying tunnel at 100\u0026thinsp;\u0026plusmn;\u0026thinsp;20\u0026deg;C. Right after drying, \u003cem\u003eF. vesiculosus\u003c/em\u003e was ground and stored in opaque plastic buckets at room temperature for further experiments.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Enzyme-Assisted Extraction (EAE) and Water Extraction (WE) of \u003cem\u003eFucus vesiculosus\u003c/em\u003e Linnaeus\u003c/h2\u003e \u003cp\u003eA proximate analysis was conducted to aid selection of appropriate enzymes for seaweed hydrolysis. Seventeen commercial enzymatic preparations were separately evaluated for the extraction of \u003cem\u003eFucus vesiculosus\u003c/em\u003e. The operational pH and temperature were chosen according to the suppliers\u0026rsquo; recommendations (data not shown). Three commercial enzymatic preparations were thus selected \u0026ndash; two glycosidases, G1 and G3, and one protease, P2 (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) with respective activities based on the supplier's datasheets.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCharacteristics of the enzymatic preparations used for extraction of \u003cem\u003eFucus vesiculosus.\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eEnzyme\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eActivity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eSource\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eForm\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eOperational pH\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eOperational\u003c/p\u003e \u003cp\u003etemperature\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGlycosidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eG1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eβ-glucosidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eAspergillus niger\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePowder\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50\u0026ndash;65\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eG3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eβ-1,3-glucanase and \u003cem\u003eBotrytis\u003c/em\u003e glucanase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eTrichoderma longibrachiatum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLiquid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProtease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eP2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eProtease (neutral) (endo)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eBacillus amyloliquefaciens\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePowder\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50\u0026deg;C\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eEAE was performed as described by Hardouin et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), with minor modifications. The seaweed powder (2.5% solid/liquid ratio (w/w)) was added to 1 L of distilled water in a jacketed glass reactor vessel. The pH was adjusted for each enzyme using either hydrochloric acid (HCl) or sodium hydroxide (NaOH). Enzymes added at a concentration of 5% (enzyme/seaweed ratio (w/w)) were placed in a 50\u0026deg;C water bath with 150 rpm of continuous tangential agitation. After 17 h, the enzymes were denatured by raising the temperature to 90\u0026deg;C for 15 min. Water extracts (WE) were obtained from seaweed powder processed in the same conditions (including the denaturation step) without enzyme addition. The extracts (WE and EAE) were centrifuged (5000 g for 15 min at 20\u0026deg;C) to remove the undigested residue, and then freeze-dried. Each extraction, WE and EAE, was performed in triplicate. The dry matter extraction yield (EY) was obtained by calculating the ratio between the total weight of dry extract (m\u003csub\u003e1\u003c/sub\u003e) and the total weight of dry matter in the reactor (m\u003csub\u003e2\u003c/sub\u003e), and results are expressed in percentage (Eq.\u0026nbsp;\u003cspan refid=\"Equ1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equ1\" name=\"EquationSource\"\u003e\n$$\\:EY=\\:\\frac{m1\\left(g\\right)}{m2\\:\\left(g\\right)}\\:x\\:100\\%\\:$$\u003c/div\u003e\u003cdiv class=\"EquationNumber\"\u003e1\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Analysis of biochemical composition\u003c/h2\u003e \u003cp\u003eAnalyses were carried out on seaweed powder and on extracts (EAE and WE). Firstly, for some analyzes (total neutral sugars and proteins content), the \u003cem\u003eF. vesiculosus\u003c/em\u003e powder was subjected to acid hydrolysis to improve their recovery and quantification according to Hardouin et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Thus, 5 mL of 1 M HCl were added to 10 mg of dry seaweed and incubated for 2 h at 100\u0026deg;C in a dry bath. Then, the solution was neutralized by adding 5 mL of 1 M NaOH. All chemical analyses were performed in triplicate (N\u0026thinsp;=\u0026thinsp;3).\u003c/p\u003e \u003cp\u003eSecondly, for other analyses (including total phenolic content and antioxidant activity), a multistep extraction process was conducted on \u003cem\u003eF. vesiculosus\u003c/em\u003e powder. Initially, 5 mg of \u003cem\u003eF. vesiculosus\u003c/em\u003e powder was mixed with 1 mL of distilled water and incubated for 2 hours at 100\u0026deg;C in a thermoshaker set to 1000 rpm. The resulting supernatant was then collected. The remaining precipitate underwent a second extraction with the addition of 1 mL of 1 M HCl, followed by another 2-hour incubation under the same conditions. The supernatant from this second extraction was combined with the first and neutralized using 1 mL of 1 M NaOH. Subsequently, the precipitate was resuspended in 1 mL of 5 M NaOH and incubated again. The final supernatant was collected and neutralized with 1 mL of 5 M HCl. All chemical analyses were performed in triplicate (n\u0026thinsp;=\u0026thinsp;3).\u003c/p\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.2.1 Total sugars\u003c/h2\u003e \u003cp\u003eAnalysis of total sugars content was performed according to the method described by Dubois et al. (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1956\u003c/span\u003e). 1 mL of tested sample or standard (glucose) (Avocado Research Chemicals Limited) was inserted into a hemolysis tube. 15 \u0026micro;L of 75% phenol (Prolabo) were added. After stirring the solution, 2.5 mL of 96% H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e were added to the tube. The solution was cooled in an ice bath for 10 min and then placed in a water bath at 30\u0026deg;C for 10 min. After stirring, the solutions were ultrasonicated for 5 s to remove micro air bubbles. The absorbance was read at 490 nm with a spectrophotometer UV-1800 (Shimadzu) and the results are expressed in percentage of dry seaweed (d.s.) or percentage of dry extract (d.e.). The standard glucose range was performed at 20, 40, 60, 80 and 100 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.2.2 Reducing sugars\u003c/h2\u003e \u003cp\u003eThis analysis was carried out on extracts (WE and EAE) according to the Miller method (1959). First, a solution of 3,5-dinitrosalicylic acid (DNS) was prepared by dissolving 1 g of DNS in 70 mL of 0.8 M NaOH while heating. After that, 30 g of sodium potassium tartrate were added before making up the solution to 100 mL with distilled water, which was then kept in the dark. 500 \u0026micro;L of the sample or standard solution (glucose) (Avocado Research Chemicals Limited) were added to 500 \u0026micro;L DNS reagent in Eppendorf tubes. The sample solutions were heated in a water bath for 10 min at 95\u0026deg;C and cooled immediately in an ice bath for 5 min. Then, the samples were incubated 15 min in the dark before taking a 200 \u0026micro;L aliquot and measuring its absorbance at 540 nm on a 96-well plate using a Multiskan GO plate reader (Thermo Scientific). The results are expressed in percentage of dry mass of extract (d.e.). The glucose concentrations were determined using a range of standard solutions at 50, 100, 150, 200, 250, 300, 400 and 500 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.2.3 Uronic acids\u003c/h2\u003e \u003cp\u003eThe composition of uronic acids was analyzed using the colorimetric method originally developed by Blumenkrantz and Asboe-Hansen in 1973, and later modified by Filisetti-Cozzy and Carpita in 1991. When exposed to hot concentrated strong acid, uronic acids dehydrate and then cyclize, forming derivatives of 5-formylfuroic acid. These derivatives react with meta-hydroxydiphenyl (MHDP) to produce a pink chromophore that absorbs light at λ\u0026thinsp;=\u0026thinsp;525 nm. In addition, the coloration is brighter in the presence of borate, but remains very sensitive to interference from neutral sugars, which stain in the presence of concentrated sulphuric acid. For this reason, potassium sulfamate is added to limit these interferences (Pierre \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Two hundred \u0026micro;L of the tested sample or standard (glucuronic acid from Acros Organics) were placed into a hemolysis tube. Subsequently, twenty \u0026micro;L of 4 M sulfamic acid (Alfa Aesar) were added. After stirring the mixture, 1.2 mL of a 75 M sodium tetraborate solution (from Merck) was carefully added. The tubes were then capped and incubated at 80\u0026deg;C for 20 minutes. Following incubation, the tubes were cooled in ice for 5 minutes. Forty \u0026micro;L of a 0.15% aqueous solution of meta-hydroxydiphenyl (MHDP) (Acros Organics) were then added. The reaction was allowed to proceed for 10 minutes before measuring the optical density at 525 nm using a Shimadzu UV-1800 spectrophotometer. The glucuronic acid standard curve was prepared with concentrations of 20, 40, 60, 80, and 100 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The results were expressed as mg/g of dry mass of seaweed (d.s.) or as a percentage of dry mass of extract (d.e.). The concentration and content of uronic acids were calculated in the same manner as for total sugars.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.2.4 Proteins\u003c/h2\u003e \u003cp\u003eThe total amount of proteins was determined using the colorimetric method developed by Smith et al. (\u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e1985\u003c/span\u003e). 25 \u0026micro;L of sample or standard solution were placed in a 96-well flat-bottom microplate, to which was added 200 \u0026micro;L of the kit reagent (Pierce BCA Protein Assay Kit, Thermo Scientific). The microplate was incubated at 37\u0026deg;C for 30 min before reading at 562 nm on a Thermo Scientific\u0026trade; Multiskan\u0026trade; GO Microplate Spectrophotometer. Bovine Serum Albumin (BSA) was used as the standard, with concentrations of 50, 100, 250, 500, 1000 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The results were expressed in percentage of dry mass of seaweed (d.s.) or percentage of dry mass of extract (d.e.).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e2.2.5 Ash\u003c/h2\u003e \u003cp\u003eThe mineral matter content was determined for \u003cem\u003eF. vesiculosus\u003c/em\u003e powder according to the method of Hardouin et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The sample was placed in a crucible and calcined by ignition. The ash content was determined thermogravimetrically after calcination of 100 mg of seaweed powder followed by a passage for 2 h in a Carbolite CSF Muffle Furnace (UK) at 585\u0026deg;C. The final mass corresponds to the mineral matter in the sample and was expressed in percentage of dry mass of seaweed (d.s.).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e2.2.6 Multi-elemental composition\u003c/h2\u003e \u003cp\u003eThe total contents of elements in \u003cem\u003eF. vesiculosus\u003c/em\u003e and extracts (WE and EAE) were determined using Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) (Agilent bench-top simultaneous optical emission Ar-ICP spectrometer (model 720)) following microwave-assisted closed-vessel digestion (Multiwave PRO microwave reaction system (Anton Paar GmbH)). For this purpose, 0.25 g of sample was placed in a PTFE (Polytetrafluoroethylene) vessel, to which were added 5 mL of 65% HNO\u003csub\u003e3\u003c/sub\u003e (Merck KGaA, Darmstadt). Then, the mixture underwent digestion for 60 min at a maximum temperature of 190\u0026deg;C. After cooling at room temperature, samples were adjusted to 25 g with deionized water and placed in a cool chamber until measurement.\u003c/p\u003e \u003cp\u003eAnalysis of nitrogen was realized using an elemental FLASH 2000 CHNS analyzer (Thermo Fisher Scientific, Waltham, MA, United States). Briefly, 2.5 mg of the sample were put in an Universal Soft Tin Containers (OD: 5 mm; H\u0026thinsp;=\u0026thinsp;8 mm, V\u0026thinsp;=\u0026thinsp;157 \u0026micro;L from Thermo Fisher Scientific, Waltham, MA, United States). Calibration was released using a standard of aspartic acid (Merck, Darmstadt, Germany). Analysis was conducted with the instruction of the manufacturer.\u003c/p\u003e \u003cp\u003eThe results were expressed as mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of dry mass of seaweed (d.s.) or mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of dry mass of extract (d.e.). Analyses were performed in duplicate.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e \u003ch2\u003e2.2.7 Plant Growth Regulators (PGRs)\u003c/h2\u003e \u003cp\u003eThe analysis of Plant Growth Regulators was performed on \u003cem\u003eF. vesiculosus\u003c/em\u003e raw material without acidic pretreatment, and extracts (EAE and WE). Methods for the quantification of Plant Growth Regulators have been previously described (Haddad et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; R\u0026eacute;thor\u0026eacute; et al. \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). 20 mg of sample were suspended in H\u003csub\u003e2\u003c/sub\u003eO (Milli-Q, 18.2 MΩ\u0026middot;cm, Millipore) containing isotope-labelled internal standards. The extracts were then purified by SPE (Solid phase extraction) using Evolute express CX (cation exchange), Biotage express plates and ABN columns (Acidic, Basic and Neutral) for cytokinins and other PGRs according to the manufacturer\u0026rsquo;s protocol.\u003c/p\u003e \u003cp\u003eThe analyses of PGRs were performed by LC-MS using a Nexera X2 UHPLC system (Shimadzu) coupled to a QTrap 6500\u0026thinsp;+\u0026thinsp;mass spectrometer (Sciex). The separation was carried out by injecting a 2 \u0026micro;L sample into a Kinetex Evo C18 core\u0026ndash;shell column (100 \u0026times; 2.1 mm, 2.6 \u0026micro;m, Phenomenex) at a flow rate of 0.7 mL min\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, with a column temperature maintained at 40\u0026deg;C. The mobile phases were composed of solvent A Milli-Q water containing 0.1% formic acid, and solvent B acetonitrile LCMS grade containing 0.1% formic acid. The analyses were performed in scheduled MRM mode. An external calibration curve was established for each PGR. A quality control sample containing all external and internal standards was injected every 10 samples to assess the system stability. The calculated concentration was corrected by the internal standard recovery rate. The results were expressed in \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of dry mass of seaweed (d.s.) or dry mass of extract (d.e.).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e \u003ch2\u003e2.2.8 Total phenolic content (TPC)\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eFolin-Ciocalteu method is commonly used for measuring the total phenolic content (TPC). This assay is based on the reduction-oxidation (redox) reactions, which are usually considered to be relatively stoichiometric and on the redox potential of the phenolic hydroxyl group (Hagerman and Butler \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). This analysis was performed directly on \u003cem\u003eF. vesiculosus\u003c/em\u003e raw material, on extracts after EAE and on WE from \u003cem\u003eF. vesiculosus\u003c/em\u003e. Briefly, in a 96-well microplate, 20 \u0026micro;L of samples (1 mg mL\u003csup\u003e-1\u003c/sup\u003e) or standard (Phloroglucinol from Sigma Aldrich) was added to 100 \u0026micro;L of Folin-Ciocalteu reagent (2N) followed by an incubation of 5 min at 30\u0026deg;C. Then, 80 \u0026micro;L of Na\u003csub\u003e2\u003c/sub\u003eCO\u003csub\u003e3\u003c/sub\u003e solution (7.5%; m/v) was added. The microplate was incubated in the dark for 30 mins min at 40\u0026deg;C. Optical density was measured at 760 nm (Waterhouse 2003; Singleton 1999). Phloroglucinol was used as standard prepared in different concentrations ranging from 0 to 1000 \u0026micro;g mL\u003csup\u003e-1\u003c/sup\u003e. The results were expressed in milligrams of Phloroglucinol Equivalent (PE) per g of dry mass of seaweed (d.s.) or dry mass of extract (d.e.).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Evaluation of antioxidant activities - DPPH Free radical scavenging assay\u003c/h2\u003e \u003cp\u003eDPPH with stable groups is a typical reagent for evaluating antioxidant activity and can be used as a representative to appraise the scavenging activity of antioxidant compounds against free radicals. This analysis is based on modified method of Guo and Liu (2012). A series of ascorbic acid solutions, served as control, was prepared in different concentrations (0\u0026ndash;30 \u0026micro;g mL\u003csup\u003e-1\u003c/sup\u003e). 100 \u0026micro;L of 0.25 mM DPPH solution were introduced to 100 \u0026micro;L sample solution or control on a 96-well microplate. Sample solutions from EAE and WE extracts were prepared in different concentrations by diluting the stock solution in methanol (0-1000 \u0026micro;g mL\u003csup\u003e-1\u003c/sup\u003e). Before reading the optical density at 517 nm, all samples were incubated at 40\u0026deg;C for 30 min in the dark. Percentage of inhibition was calculated using the following formula (2):\u003cdiv id=\"Equ2\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equ2\" name=\"EquationSource\"\u003e\n$$\\:I\\:\\left(\\%\\right)=\\:{\\left[\\right(A}_{C}-{A}_{S})/{A}_{C}]\\:x\\:100$$\u003c/div\u003e\u003cdiv class=\"EquationNumber\"\u003e2\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003ewhere: I (%)\u0026thinsp;=\u0026thinsp;inhibition (expressed in %), A\u003csub\u003eC\u003c/sub\u003e = absorbance of control, and A\u003csub\u003eS\u003c/sub\u003e = absorbance of samples. IC\u003csub\u003e50\u003c/sub\u003e, corresponding to the concentration sufficient to obtain 50% of a maximum scavenging capacity of samples was determined based on the regression obtained from dose\u0026ndash;response curve.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Statistical analyses\u003c/h2\u003e \u003cp\u003eThe statistical analyses were carried out using R software and RStudio interface, 2023.09.01 version, to evaluate the statistically significant differences (for p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) between examined groups. Multiple comparison analyses were used for this comparative study between extracts obtained from \u003cem\u003eF. vesiculosus\u003c/em\u003e. They include parametric (ANOVA) and non-parametric (Kruskall-Wallis) tests after checking the normality and equal variances of data. These analyses used various packages such as rstatix, RcmdrMisc, multcomp, rcompanion and FSA (Fox et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Hollander et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1973\u003c/span\u003e; Chambers et al. 1992).\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 \u003cem\u003eFucus vesiculosus\u003c/em\u003e powder\u003c/h2\u003e\n \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e\n \u003ch2\u003e3.1.1 Proximate composition\u003c/h2\u003e\n \u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e presents the biochemical composition of the \u003cem\u003eF. vesiculosus\u003c/em\u003e raw material. Although \u003cem\u003eFucus vesiculosus\u003c/em\u003e is a very common seaweed species in the Brittany coast, little research has been conducted on the study of its biochemical composition.\u003c/p\u003e\n \u003cp\u003eAccording to the results, \u003cem\u003eF. vesiculosus\u003c/em\u003e is mainly composed of neutral sugars which constitute 22% of d.s. Proteins are the second important component found in \u003cem\u003eF. vesiculosus\u003c/em\u003e which constitute 17% of d.s. The uronic acids constitute 12% of d.s. however there is no information was found in the literature concerning the content of uronic acids in \u003cem\u003eFucus vesiculosus\u003c/em\u003e biomass. In addition, the raw \u003cem\u003eF. vesiculosus\u003c/em\u003e presents 39.8\u0026thinsp;\u0026plusmn;\u0026thinsp;9.8 mg eq PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s. of total phenolic content which consists of approximately 4% of dry weight.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec18\" class=\"Section3\"\u003e\n \u003ch2\u003e3.1.2 Multi-element analysis\u003c/h2\u003e\n \u003cp\u003eMulti-element composition of \u003cem\u003eFucus vesiculosus\u003c/em\u003e is presented in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. Elements are classified as macroelements (essential for plant growth and development) and microelements (necessary for a number of physiological functions). The three main elements, in term of concentration, present in \u003cem\u003eF. vesiculosus\u003c/em\u003e are potassium and sodium with 25 993 mg Kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s. and 25 497mg Kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s., respectively and calcium with 15 966 mg Kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eMulti-element composition of \u003cem\u003eFucus vesiculosus\u003c/em\u003e (mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s.) (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD; n\u0026thinsp;=\u0026thinsp;2)\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"2\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eElement\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eF. vesiculosus powder material\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMacroelements\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePotassium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e25 993\u0026thinsp;\u0026plusmn;\u0026thinsp;312\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSodium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e25 497\u0026thinsp;\u0026plusmn;\u0026thinsp;312\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCalcium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15 966\u0026thinsp;\u0026plusmn;\u0026thinsp;335\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSulphur\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10 818\u0026thinsp;\u0026plusmn;\u0026thinsp;834\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNitrogen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8 831\u0026thinsp;\u0026plusmn;\u0026thinsp;762\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMagnesium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8 603\u0026thinsp;\u0026plusmn;\u0026thinsp;155\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePhosphorus\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e716\u0026thinsp;\u0026plusmn;\u0026thinsp;9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicroelements\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIron\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e569\u0026thinsp;\u0026plusmn;\u0026thinsp;11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBoron\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e118\u0026thinsp;\u0026plusmn;\u0026thinsp;3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eManganese\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e106\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eZinc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCopper\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\n \u003ch2\u003e3.1.3 Plant Growth Regulators\u003c/h2\u003e\n \u003cp\u003eTable \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e represents the content of Plant Growth Regulators in in\u0026nbsp;\u003cem\u003eFucus vesiculosus\u003c/em\u003e.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eContent of Plant Growth Regulators in \u003cem\u003eFucus vesiculosus\u003c/em\u003e (\u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s.; mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD; n\u0026thinsp;=\u0026thinsp;3).\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"2\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlant Growth Regulators\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eContent in F. vesiculosus\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSalicylic acid (SA)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e113\u0026thinsp;\u0026plusmn;\u0026thinsp;3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAuxin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIndole-3-acetic acid (IAA)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e54\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e1-amino-1-cyclopropane-carboxylic acid (ACC)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e37\u0026thinsp;\u0026plusmn;\u0026thinsp;3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eDihydrojasmonic acid (DJA)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAbscisic acid (ABA)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eCytokinins\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCis zeatin (cZ)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIsopentenyladenine (iP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIsopentenyladenosine (iPR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTran zeatin (tZ)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eAccording to Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, three PGRs are most represented in \u003cem\u003eF. vesiculosus\u003c/em\u003e. The salicylic acid was detected in large quantity with up to 112.7 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of d.s., followed by auxin (53.5 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s.) and the ethylene precursor \u0026ndash; ACC (1-amino-cyclopropane-1-carboxylic acid) (36.9 \u0026micro;g/kg d.s.).\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 \u003cem\u003eFucus vesiculosus-\u003c/em\u003ederived extracts\u003c/h2\u003e\n \u003cdiv id=\"Sec21\" class=\"Section3\"\u003e\n \u003ch2\u003e3.2.1 Biochemical composition\u003c/h2\u003e\n \u003cp\u003eFigure 2 presents the dry matter extraction yield (Figure 2(a)) and the biochemical composition of the extracts (Figure 2 (b-f)) obtained from\u0026nbsp;\u003cem\u003eF. vesiculosus\u003c/em\u003e after EAE and WE, according to the operational pH of each enzyme (one protease P2 at pH 7, two different glucosidases G1 and G3 at respectively pH 4 and pH 5). \u0026nbsp;\u003c/p\u003e\n \u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. (a) Extraction yield expressed as the mass of dry matter extracted per 100 g of dry \u003cem\u003eF. vesiculosus\u003c/em\u003e and biochemical composition of \u003cem\u003eFucus\u003c/em\u003e-derived extracts expressed in percentage of dry extract (d.e.) \u0026ndash; (b) total sugars; (c) reducing sugars; (d) uronic acids (% of dry mass of extract (d.e.); (e) proteins; (e); (f) Total Phenolic Content (mg eq phloroglucinol PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.) mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD; n\u0026thinsp;=\u0026thinsp;3.\u003c/p\u003e\n \u003cp\u003eResults obtained for the dry matter extraction yield (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(a)) showed that pH influences the recovery of compounds from \u003cem\u003eF. vesiculosus\u003c/em\u003e. Significantly higher dry matter extraction yields were observed with WE at pH 4 and pH 7, showing an increase of 18% (p\u0026thinsp;=\u0026thinsp;0.001) and 14% (p\u0026thinsp;=\u0026thinsp;0.003) respectively, compared to WE at pH 5. Notably, only G3 enzyme enhanced the dry matter extraction yield, presenting a statistically significant increase of 13% (p\u0026thinsp;=\u0026thinsp;0.006) over WE at pH 5. Among the enzymes tested, G1 achieved the highest dry matter extraction yield at 60%, outperforming both enzyme G3 (p\u0026thinsp;=\u0026thinsp;0.026) and P1 (p\u0026thinsp;=\u0026thinsp;0.024).\u003c/p\u003e\n \u003cp\u003eRegarding total sugars (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(b)), only the enzyme G3 allowed statistical increase (+\u0026thinsp;34%) in the extraction of these compounds in comparison with the corresponding WE at pH 5. The extract obtained with G3 contained the highest amount of total sugars (38\u0026thinsp;\u0026plusmn;\u0026thinsp;4% of d.e.) in comparison with the two other enzymes G1 and P2 with statistically significant differences between these groups (i.e., G1 and G3 and P2) with results multiplied by 1.8 (\u003cem\u003ep\u003c/em\u003e 0.0008) and by 2 (\u003cem\u003ep\u003c/em\u003e 0.0004) respectively).\u003c/p\u003e\n \u003cp\u003eFor both glycosidases (G1 and G3) significant higher amounts of reducing sugars were obtained (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(c)) equal to 21% and 10% d.e., respectively with significant differences in comparison with WE at pH 4 and WE and pH 5. Finally, even if extracts obtained with G1 had lower content of total sugars in comparison with G3, they contained twice more reducing sugars in comparison with G3 (\u003cem\u003ep\u003c/em\u003e 0.0002).\u003c/p\u003e\n \u003cp\u003eThe uronic acid content (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(d)) varied between 7% and 10% d.e., with no significant differences among the different extracts, except for those with P2 at pH 7. These latter extracts had the lowest uronic acid content at 6.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4% d.e., while the WE pH 5 extracts had the highest at 9.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8% d.e.\u003c/p\u003e\n \u003cp\u003eRegarding proteins, the conventional water extracts contained 39\u0026thinsp;\u0026plusmn;\u0026thinsp;3% d.e. of proteins at pH 5, and this increased to 53\u0026thinsp;\u0026plusmn;\u0026thinsp;3% d.e. at pH 4 (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(e)). Statistically significant differences were observed between WE at pH 4 and pH 5 (p\u0026thinsp;=\u0026thinsp;0.004), and between WE at pH 7 and pH 5 (p\u0026thinsp;=\u0026thinsp;0.005). Extracts obtained with G1 and P2 had higher protein content compared to G3, with statistically significant differences.\u003c/p\u003e\n \u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(f) shows the total phenolic content (TPC) of the different extracts. The results indicate that the highest TPC was found for the extracts obtained by EAE carried out with P2 enzyme at pH 7 (116\u0026thinsp;\u0026plusmn;\u0026thinsp;7 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.) with twice more TPC in comparison with its WE at pH 7 (54\u0026thinsp;\u0026plusmn;\u0026thinsp;11 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.). On the other hand, extract produced with G1 enzyme at pH 4 yielded a higher TPC (71\u0026thinsp;\u0026plusmn;\u0026thinsp;11 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.) than its WE at the same pH (58\u0026thinsp;\u0026plusmn;\u0026thinsp;11 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e). Finally, G3 enzyme extract at pH 5 showed a TPC content of 57\u0026thinsp;\u0026plusmn;\u0026thinsp;8 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e while the WE extract at pH 5 yielded a TPC content of 50\u0026thinsp;\u0026plusmn;\u0026thinsp;9 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec22\" class=\"Section3\"\u003e\n \u003ch2\u003e3.2.2 Multi-element analysis\u003c/h2\u003e\n \u003cp\u003eTable \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e summarizes the multi-element composition of extracts obtained from \u003cem\u003eFucus vesiculosus\u003c/em\u003e by EAE and WE.\u003c/p\u003e\n \u003cp\u003eWhatever the method and conditions of extraction, the most abundant elements are sodium, potassium and sulphur. It appears that conventional WE most often leads to similar or higher contents in macro- and microelements in aqueous extracts in comparison with extracts produced with EAE. Indeed, only few results show that enzymes allow for increasing the content of elements in the extract (d.e.), for example the content of nitrogen. The highest nitrogen content in the dry mass of the extract was obtained when P2 or G1 were used, leading to higher quantities that their respective WE at same pH.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eMulti-element composition of extracts produced from \u003cem\u003eFucus vesiculosus\u003c/em\u003e by WE (pH 4 and pH 5) or EAE (enzymes G1, G3, P2) (mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.; mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD; n\u0026thinsp;=\u0026thinsp;2)\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eElement\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWE pH 4\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWE pH 5\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWE pH 7\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eG1 pH 4\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eG3 pH 5\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP2 pH7\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMacroelements\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSodium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e43 852\u0026thinsp;\u0026plusmn;\u0026thinsp;614\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42 913\u0026thinsp;\u0026plusmn;\u0026thinsp;996\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e44 225\u0026thinsp;\u0026plusmn;\u0026thinsp;388\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e47 397\u0026thinsp;\u0026plusmn;\u0026thinsp;538\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e39 635\u0026thinsp;\u0026plusmn;\u0026thinsp;692\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e45 035\u0026thinsp;\u0026plusmn;\u0026thinsp;676\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePotassium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e40 623\u0026thinsp;\u0026plusmn;\u0026thinsp;325\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e39 643\u0026thinsp;\u0026plusmn;\u0026thinsp;919\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e38 603\u0026thinsp;\u0026plusmn;\u0026thinsp;46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e37 861\u0026thinsp;\u0026plusmn;\u0026thinsp;644\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e36 081\u0026thinsp;\u0026plusmn;\u0026thinsp;589\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e39 005\u0026thinsp;\u0026plusmn;\u0026thinsp;702\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSulphur\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14 128\u0026thinsp;\u0026plusmn;\u0026thinsp;48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14 465\u0026thinsp;\u0026plusmn;\u0026thinsp;248\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13 676\u0026thinsp;\u0026plusmn;\u0026thinsp;11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13 534\u0026thinsp;\u0026plusmn;\u0026thinsp;257\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12 476\u0026thinsp;\u0026plusmn;\u0026thinsp;73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12 627\u0026thinsp;\u0026plusmn;\u0026thinsp;278\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMagnesium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7 988\u0026thinsp;\u0026plusmn;\u0026thinsp;19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7 223\u0026thinsp;\u0026plusmn;\u0026thinsp;76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6 940\u0026thinsp;\u0026plusmn;\u0026thinsp;38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7 544\u0026thinsp;\u0026plusmn;\u0026thinsp;106\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6 653\u0026thinsp;\u0026plusmn;\u0026thinsp;65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6 694\u0026thinsp;\u0026plusmn;\u0026thinsp;74\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNitrogen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4 878\u0026thinsp;\u0026plusmn;\u0026thinsp;470\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5 659\u0026thinsp;\u0026plusmn;\u0026thinsp;40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5 330\u0026thinsp;\u0026plusmn;\u0026thinsp;320\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5 844\u0026thinsp;\u0026plusmn;\u0026thinsp;210\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5 767\u0026thinsp;\u0026plusmn;\u0026thinsp;744\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6 188\u0026thinsp;\u0026plusmn;\u0026thinsp;288\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCalcium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7 097\u0026thinsp;\u0026plusmn;\u0026thinsp;38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4 980\u0026thinsp;\u0026plusmn;\u0026thinsp;42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4 962\u0026thinsp;\u0026plusmn;\u0026thinsp;42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6 469\u0026thinsp;\u0026plusmn;\u0026thinsp;45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4 962\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4 805\u0026thinsp;\u0026plusmn;\u0026thinsp;30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePhosphorus\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e644\u0026thinsp;\u0026plusmn;\u0026thinsp;8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e741\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e782\u0026thinsp;\u0026plusmn;\u0026thinsp;16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e699\u0026thinsp;\u0026plusmn;\u0026thinsp;6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e752\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e753\u0026thinsp;\u0026plusmn;\u0026thinsp;9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicroelements\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBoron\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e180\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e190\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e192\u0026thinsp;\u0026plusmn;\u0026thinsp;5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e168\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e181\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e181\u0026thinsp;\u0026plusmn;\u0026thinsp;3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIron\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e184\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e139\u0026thinsp;\u0026plusmn;\u0026thinsp;5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e101\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e179\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e162\u0026thinsp;\u0026plusmn;\u0026thinsp;4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e96\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eManganese\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e91\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e86\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e83\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e87\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e83\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e76\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eZinc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e22\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e23\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e23\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCopper\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e\n \u003ch2\u003e3.2.3 Plant Growth Regulators\u003c/h2\u003e\n \u003cp\u003eTable \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e presents the content of PGRs in extracts obtained through WE and EAE. As observed for \u003cem\u003eF. vesiculosus\u003c/em\u003e powder (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e), the three main PGRs present in \u003cem\u003eF. vesiculosus\u003c/em\u003e extracts were SA, IAA and ACC. Regarding the comparison of PGRs detected in raw \u003cem\u003eF. vesiculosus\u003c/em\u003e powder and extracts, Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e reveals that new PGRs were identified in the extracts, such as cZR and tZR which are precursors of cZ and tZ respectively. Conversely, DJA is not anymore detected in the extracts.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003ePlant Growth Regulators composition of the extracts produced from \u003cem\u003eFucus vesiculosus\u003c/em\u003e (\u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.; mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD; n\u0026thinsp;=\u0026thinsp;3)\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"22\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlant growth regulators\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWE pH 4\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eWE pH 5\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWE pH 7\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eG1 pH 4\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eG3 pH 5\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eP2 pH7\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAuxin\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eIndole-3-acetic acid (IAA)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e62\u0026thinsp;\u0026plusmn;\u0026thinsp;6\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e88\u0026thinsp;\u0026plusmn;\u0026thinsp;4\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e325\u0026thinsp;\u0026plusmn;\u0026thinsp;34\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e58\u0026thinsp;\u0026plusmn;\u0026thinsp;4\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e105\u0026thinsp;\u0026plusmn;\u0026thinsp;12\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e289\u0026thinsp;\u0026plusmn;\u0026thinsp;35\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSalicylic acid (SA)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e213\u0026thinsp;\u0026plusmn;\u0026thinsp;8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e245\u0026thinsp;\u0026plusmn;\u0026thinsp;17\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e289\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e177\u0026thinsp;\u0026plusmn;\u0026thinsp;3\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e245\u0026thinsp;\u0026plusmn;\u0026thinsp;10\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e297\u0026thinsp;\u0026plusmn;\u0026thinsp;5\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e1-amino-1-cyclopropane-carboxylic acid (ACC)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e86\u0026thinsp;\u0026plusmn;\u0026thinsp;35\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e103\u0026thinsp;\u0026plusmn;\u0026thinsp;4\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e255\u0026thinsp;\u0026plusmn;\u0026thinsp;29\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e57\u0026thinsp;\u0026plusmn;\u0026thinsp;10\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e165\u0026thinsp;\u0026plusmn;\u0026thinsp;45\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e207\u0026thinsp;\u0026plusmn;\u0026thinsp;22\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAbscisic acid (ABA)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e3\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e4\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eCytokinins\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIsopentenyladenosin (iPR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e12\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e4\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eIsopentenyladenine (iP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCis zeatin (cZ)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e28\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTran zeatin riboside (tZR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e3\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCis zeatin riboside (cZR)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTran zeatin (tZ)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"7\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec24\" class=\"Section3\"\u003e\n \u003ch2\u003e3.2.4. Biological anti-oxidant activity\u003c/h2\u003e\n \u003cp\u003eThe half maximal Inhibitory Concentration IC\u003csub\u003e50\u003c/sub\u003e values resulting from the DPPH test for the various \u003cem\u003eF. vesiculosus\u003c/em\u003e powder and extracts are presented in Table \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e. The IC\u003csub\u003e50\u003c/sub\u003e values across different extracts ranged between 177.3 and 212.6 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and are not all significantly different, making challenging the comparisons between DPPH scavenging activity of EAE and WE extracts. The highest total phenolic content was observed in the protease extract at pH 7 (116.4\u0026thinsp;\u0026plusmn;\u0026thinsp;6.8 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.). Despite this, it did not exhibit the greatest antioxidant activity. Surprisingly, it was the WE at pH 4 that showed the highest antioxidant activity, despite having a lower total phenolic content (57.7\u0026thinsp;\u0026plusmn;\u0026thinsp;11 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e.) with an IC\u003csub\u003e50\u003c/sub\u003e value of 177.3 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab6\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eDry weight rate and antioxidant activity (IC\u003csub\u003e50\u003c/sub\u003e) of the \u003cem\u003eFucus vesiculosus\u003c/em\u003e powder and extracts\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"14\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003en\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eDry weight rate\u003c/p\u003e\n \u003cp\u003e(% f.w.)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eTotal phenolic content\u003c/p\u003e\n \u003cp\u003e(mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e or d.s)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eAntioxidant activity\u003c/p\u003e\n \u003cp\u003e(IC\u003csub\u003e50\u003c/sub\u003e \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026plusmn;\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026plusmn;\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e\u0026plusmn;\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e**\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003eF. vesiculosus\u003c/em\u003e powder\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e91.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026plusmn;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ed\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e39.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026plusmn;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e309.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026plusmn;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e33.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003ec\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWE pH 4 extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eabc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e177.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWE pH 5 extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eab\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e189.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWE pH 7 extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e54.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eabc\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e212.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eb\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eP2 pH 7 extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e116.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ed\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e188.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eG1 pH 4 extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eb\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e70.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e195.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eab\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eG3 pH 5 extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ec\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e56.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eacd\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e186.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cem\u003e\u0026plusmn;\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ea\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"13\"\u003e\n \u003cp\u003e* One-way analysis of means, not assuming equal variances, \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"13\"\u003e\n \u003cp\u003e** One-way analysis of means, not assuming equal variances, \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e\n \u003cp\u003eIC\u003csub\u003e50\u003c/sub\u003e of ascorbic acid (control): 15.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cdiv id=\"Sec26\" class=\"Section2\"\u003e \u003ch2\u003e4.1 Characterization of \u003cem\u003eFucus vesiculosus\u003c/em\u003e powder\u003c/h2\u003e \u003cp\u003eComparing the data obtained with existing literature is challenging. Indeed, various factors, including those related to the harvesting, as well as others such as biomass pre-treatment (such as drying and grinding processes), selection of biochemical analysis techniques, contribute to significant variations. The main parameters responsible of variations in the biochemical composition of seaweed, including \u003cem\u003eFucus vesiculosus\u003c/em\u003e, are related to the harvest season, geographical location, environmental conditions like wave exposure and intensity, salinity, light availability, and reproductive phase or intra- and inter-specific competition (Paiva et al. 2018).\u003c/p\u003e \u003cp\u003eBased on Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, 33% of the non-detected compounds could be attributed to sulfated groups linked with fucose-rich polysaccharides, such as fucans and fucoidans (Rodr\u0026iacute;guez-Jasso et al. \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2013\u003c/span\u003e, Obluchinskaya et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). \u003cem\u003eFucus\u003c/em\u003e species are known to be rich in minerals, proteins, antioxidant phloroglucinol-based phenolic compounds, and cell wall polysaccharides, including alginates and sulphated polysaccharides rich in fucose (fucoidan) (Bikovens et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Paiva et al. 2018; Obluchinskaya et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Rousseau et al. \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The main constituents of the brown seaweed fibers are three distinct polysaccharides, namely sulfated polysaccharides rich in fucose (fucoidan), alginic acid (consists of β-D-mannuronic (M) and α-L-glucuronic (G) acid units) and laminaran (Catarino et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The contents may vary between 3.4 and 25.7% d.w. for fucoidan, 8.4 and 58.8% for alginic acid and 0.6 and 7.0% for laminaran. Obluchinskaya et al. (\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) showed the amounts of monosaccharides, phlorotannins, flavonoids, and the mineral composition significantly varied in Fucus vesiculosus with reproductive phase and geographic locations in different Arctic region. Paiva et al. (2018) also discovered seasonal and geographical differences in the nutritional composition (protein, carbohydrate, lipid and fatty acids, dietary fiber) and functional activity (total phenolic and flavonoid content, antioxidant activity) of Fucus spiralis.\u003c/p\u003e \u003cp\u003eCarbohydrates are the most abundant element of Fucus spp., involving in different roles including storage, defense responses, development, cell polarity or tissues integrity (Deniaud-Bouet et al. 2017). Carbohydrates can vary from 34\u0026ndash;66% (d.w.) in \u003cem\u003eFucus vesiculosus\u003c/em\u003e (Catarino et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The 22% of neutral sugars determined in F. vesiculosus in our study is quite similar to the result presented by Bikovens et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Collected in March 2017 in Jurmala (Latvia), the total neutral sugars content of F. vesiculosus were analyzed using the same method and presented 24.6% d.w. Raw \u003cem\u003eFucus\u003c/em\u003e collected in Galicia (Spain) is rich in both soluble (10.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31%) and insoluble (48.63\u0026thinsp;\u0026plusmn;\u0026thinsp;1.05%) fractions of dietary fibers (Diaz-Rubio et al. 2009). Fucus vesiculosus from Portugal is rich in carbohydrates (56.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4g/100 g seaweed d.w.) with fibers representing the main contribution (45.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1g/100 g d.w.). \u003cem\u003eFucus vesiculosus\u003c/em\u003e stood out by its high content of insoluble fiber compared to soluble fiber (Neto et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eProteins are the second most important compound in this \u003cem\u003eF. vesiculosus\u003c/em\u003e powder with 17% of d.s. which is in accordance with the literature. Indeed, according to Fleurence (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1999\u003c/span\u003e), most of the brown seaweeds industrially exploited (Laminaria digitata, Ascophyllum nodosum, Fucus vesiculosus and Himanthalia elongata) have a protein content lower than 15% d.w. It shows little variations, from 11% (Catarino et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), to 13% d.w. (Lorenzo et al. 2017), with up to 15% d.w. (Neto et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eWithin the Fucaceae family, phlorotannins are the main phenolic compounds found in brown algae, making them a valuable source of polyphenols (Obluchinskaya et al. \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In fact, phlorotannins have been identified as the only phenolic compounds in F. vesiculosus (Catarino et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The abundance of phlorotannins in brown marine algae, as documented in the literature, ranges from 1\u0026ndash;14% of dry algal biomass (Ar Gall et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Catarino et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Maheswaria and Babu \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In our study, the total phenolic content in the raw \u003cem\u003eF. vesiculosus\u003c/em\u003e consisted of approximately 4% of dry weight (39.8\u0026thinsp;\u0026plusmn;\u0026thinsp;9.8 mg eq PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s) which is in accordance with the literature.\u003c/p\u003e \u003cp\u003eThe mineral content in seaweed is very variable (8\u0026ndash;40% d.w.). It is primarily attributed to the properties of their cell surface polysaccharides, which facilitate the retention of inorganic marine substances. In the particular case of \u003cem\u003eFucus\u003c/em\u003e spp., the ash (mineral) content may vary from 19\u0026ndash;36% d.w. (Catarino et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Obluchinskaya et al. in 2022 confirmed that the ash content of macroalgae varies significantly, depending on the species, geographic location, and reproductive phase. The authors showed averages ash values of 18.5% and 28.5% d.w. depending on reproductive phases and locations. In our study, \u003cem\u003eFucus vesiculosus\u003c/em\u003e contains low amount of minerals with only 12% of ashes (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eFucus vesiculosus\u003c/em\u003e contains several important minerals for plants such as P, K, Ca, Mg. According to Catarino et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), sodium and potassium are the major elements described in this genus. According to Balina et al. (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), F. \u003cem\u003evesiculosus\u003c/em\u003e from Jurmala, Latvia contains 21 500 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of calcium, 11 000 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of potassium, 9 300 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of magnesium, 6 300 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of sodium, 1 680 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of manganese, 1 400 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of phosphorus, 490 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of iron, 89 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of zinc and 12.7 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of copper. The amounts of iron and magnesium obtained in this study are quite similar by contrast, phosphorus, zinc, copper and manganese are much lower (by 49\u0026ndash;94% respectively). In addition, calcium is present in lower quantity in comparison with the results of literature (lower by 26%), while potassium and sodium are present in higher quantity (increase by 136% and 305% respectively). These differences could be explained by the effect related to local harvesting conditions (i.e., Spain for Balina et al. (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and France in this study) and seasonal variations (i.e., in January for Balina et al. (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and in July in this study) as reported and confirmed by Allen et al. (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In addition, content of elements in seaweeds depend on the concentrations of metal compounds in the surrounding water and they ability to bioaccumulate such compounds (Adams et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFollowing the initial discovery of auxin in marine algae by Overbeek in 1940, research attention shifted towards identifying Plant Growth Regulators (PGRs) present in seaweeds. However, conclusive identification of auxin remained uncertain until the 1970s. During the past two decades, various PGRs (auxins, cytokinins, gibberellins, abscisic acid) were reported in different seaweed extracts (Stirk and Van Staden \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe first mention of the potential presence of PGR-type compounds in \u003cem\u003eF. vesiculosus\u003c/em\u003e dates back to 1971. Biological tests carried out on Avena coleoptiles revealed the release of several indole-type compounds by the alkaline hydrolysis of various seaweeds, including \u003cem\u003eFucus vesiculosus\u003c/em\u003e (Buggeln \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1971\u003c/span\u003e). Most recently, a study determined the content of PGRs in \u003cem\u003eF. vesiculosus\u003c/em\u003e (Sharma et al. 2012). The acidic extracts obtained from \u003cem\u003eF. vesiculosus\u003c/em\u003e collected in June contained up to 46.84 \u0026micro;g per kilogram of d.s which is in the same range with the results in this study. Regarding other brown seaweed species, the profiles and contents of PGRs have been explored in \u003cem\u003eSargassum horneri\u003c/em\u003e (Li et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The content of salicylic acid was two times lower and the amount of IAA (Indole-3-acetic acid) six times lower compared with the present study. These authors have detected 3.9 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for iP (Isopentenyladenine); 2.4 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of iPR (Isopentenyladenosine) and 5.6 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e of ABA (Abscisic acid). Yalcin et al. (\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) observed that the content of phytohormones is very low for two brown seaweeds from 3.5 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for \u003cem\u003eCladostephus spongiosum\u003c/em\u003e to 15.4 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for \u003cem\u003eCystoseira barbata\u003c/em\u003e. Stirk et al. (\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) showed a content of IAA of 2.04 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for \u003cem\u003eEcklonia maxima\u003c/em\u003e and 1.7 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for \u003cem\u003eMacrocystis pyrifera\u003c/em\u003e. However, in the present work, iP, iPR, cZ and ABA contents are quite similar to the values published by these authors. Stirk et al. (\u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) have measured an amount of iP at 0.042 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for \u003cem\u003eEcklonia maxima\u003c/em\u003e and 0.035 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for \u003cem\u003eMacrocystis pyrifera\u003c/em\u003e; iPR at 0.006 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and 0.003 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e respectively and cZR was under the limit of detection for both seaweeds.\u003c/p\u003e \u003cp\u003eIn general, PGRs from seaweeds are extracted with an organic solvent (i.e., methanol) with or without sonication. Subsequent purification by SPE or affinity chromatography is usually applied and the identification and quantification of PGRs are then performed by LC-MS or GC-MS (Benitez-Gracia et al. 2020; Yalcin et al. \u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In the present study, the PGRs from \u003cem\u003eF. vesiculosus\u003c/em\u003e were purified after extraction and concentrated by C18 or cation-exchange SPE depending on the nature of the molecule of interest. The quantification was performed by UHPLC-QTRAP-MS/MS in MRM (Multiple reaction monitoring) mode to achieve high specificity and sensitivity. The combination of an advanced chromatographic separation technique with tandem mass spectrometry allows the detection of PGRs down to ng/kg. While there are a few studies that have determined the levels of PGRs in seaweed, the final quantification is evidently affected by the method of harvesting and processing the seaweed samples after collection, including whether or not the raw material is dried.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec27\" class=\"Section2\"\u003e \u003ch2\u003e4.2 Comparison of \u003cem\u003eFucus vesiculosu\u003c/em\u003es Extracts Derived from Conventional Aqueous Extraction (WE) and Enzyme-Assisted Extraction (EAE)\u003c/h2\u003e \u003cp\u003eRegarding total sugars (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e(b)), it was expected that extracts obtained with glycosidases would contain more sugars compared with protease due to the main activity dedicated to hydrolysis of osidic compounds such as oligo and polysaccharides. Results demonstrated the efficient enzymatic hydrolysis of polysaccharides contained in the seaweed thanks to G3 which allowed an increase in total sugars. As a result, the extract obtained using G3 at pH 5 was richer in sugars compared to the conventional water extraction method. Although EAE can significantly enhance the extraction of polysaccharides from brown algae by breaking down the cell wall, no studies were found in the literature specifically comparing our results obtained with the glucosidase enzyme. It is possible to assume that even if G1 enzyme breakdowns more sugars molecules, such as polysaccharides, resulting in more reducing ends (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e (c)), the obtained polysaccharides are not sufficiently hydrolyzed to be quantified by the colorimetric method used here for the determination of total sugars. Seaweed-derived polysaccharides have the potential to act as plant biostimulants. Mzibra et al. (\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) showed that polysaccharide enriched extract obtained from \u003cem\u003eFucus spiralis\u003c/em\u003e significantly increased germination percentage, germination speed, reduced mean germination time of tomato seeds treated with this extract at a dose of 0.02 mg/mL. Moreover, the same extract applied to the soil and as a foliar spray enhanced tomato growth parameters such as leaves number, plant length, dry weight of shoot and root, and chlorophyll content. Polysaccharide enriched extract from \u003cem\u003eF. spiralis\u003c/em\u003e applied to the soil improved plant growth parameters more successfully than applied as a foliar spray.\u003c/p\u003e \u003cp\u003eRegarding proteins (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e(e)), None of the tested enzymes allowed the recovery of more proteins in comparison with their respective WE, even for the protease P2. Agregan et al. (2017) have analyzed the protein content of \u003cem\u003eF. vesiculosus\u003c/em\u003e water extracts obtained through UAE. They obtained results higher than those presented in this study with 62.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3% d.e.\u003c/p\u003e \u003cp\u003eA significant difference between the TPC (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e (f)) was found for the EAE extraction with P2 enzyme compared with WE at the different tested pH. Among these extractions, the EAE with P2 enzyme at pH 7 seems to be the most efficient for enriching total phenolic content in \u003cem\u003eF. vesiculosus\u003c/em\u003e extracts. Our findings align with the existing literature data. In fact, several studies have demonstrated that EAE can enhance the extraction efficiency of polyphenols from brown algae (Hardouin et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Teixeira-Guedes et al. \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Specifically, the disruption of the algal cell wall during EAE leads to the release of intracellular constituents, including proteins, which are prone to complex with polyphenols, causing aggregation and eventual precipitation (Siriwardhana et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). These polyphenols may interact with proteins through various mechanisms such as hydrogen bonding, π-bonding, hydrophobic interactions, as well as ionic and covalent linkages. Wang et al. (2020) investigated the application of proteases to enhance the yield of polyphenols and antioxidants from the red seaweed \u003cem\u003ePalmaria palmata\u003c/em\u003e. They observed an increase in the extraction efficiency of phenolic compounds following protease treatment, attributing this improvement to the conversion of proteins into small peptides and free amino acids. Indeed, the protease degradation of proteins can liberate bound polyphenols, thereby improving access to polyphenols and facilitating their extraction (Wang et al. 2020).\u003c/p\u003e \u003cp\u003eAccording to Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, Enzyme G1 increases phosphorus contents in enzymatic extracts in comparison with WE at pH 4 (increase by 9% respectively). The enzyme G3 permitted to increase the extraction of iron (increase by 17%) and copper (by 45%) in comparison with WE at pH 5. The comparison of the three enzymes shows that G1 allowed for improved extraction of calcium, iron, magnesium, manganese, sodium and sulphur reaching the highest content in the dry extract in comparison with the extraction with G3 and P2. On the other hand, enzyme G3 permitted to obtain the highest extraction of boron, copper, phosphorus and zinc in comparison with enzymes G1 and P2. P2 permitted to obtain the highest content of potassium and nitrogen in the extract in comparison with G1 and G3 and had an equivalent amount of phosphorus in the extract as for the enzyme G3. Finally, considering the three main macroelements mandatory for plant nutrition and growth (nitrogen, phosphorus and potassium), extracts obtained with P2 are promising with the highest amount of nitrogen 6188\u0026thinsp;\u0026plusmn;\u0026thinsp;288 mg kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e., which can be correlated to the use of a protease.\u003c/p\u003e \u003cp\u003ePlants require specific essential minerals crucial for their structural integrity and metabolic processes. Inadequate supply of these elements can result in severe abnormalities in plant growth and development. Among the macroelements, nitrogen, phosphorus, and potassium are primary nutrients found in both mineral and organic fertilizers. Nitrogen deficiency adversely impacts plant productivity by reducing photosynthesis, leaf area, and the lifespan of green foliage. Phosphorus deficiency limits plant growth and crop yield, while potassium deficiency makes plants more susceptible to stress, leading to stomatal closure and inhibition of photosynthetic rates in various crop species (Wissuwa et al. 2003; Wang et al. \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Mu et al. 2021). The current study demonstrates that EAE extracts contain a range of elements, including nitrogen, phosphorus, and potassium. Therefore, application of these extracts on the crops could supply essential minerals for healthy growth (Choulot et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Furthermore, these elements may play a role in activating plant defense mechanisms (Shukla et al. 2016). Literature data indicate that extract obtained from \u003cem\u003eF. vesiculosus\u003c/em\u003e by UAE, rich in mineral components, may have a positive effect on plant growth, increasing the plant length, weight of plants, and chlorophyll content, in comparison with the control group (Krautforst et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Michalak and Baśladyńska (\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) showed that the application of 5% \u003cem\u003eF. vesiculosus\u003c/em\u003e extract, produced by hydrolysis with potassium hydroxide, on garden cress improved not only plant length and chlorophyll content, but also biofortified cultivated plants in Cu, Fe, Mn, Zn, Ca, S, P and K.\u003c/p\u003e \u003cp\u003eThe increases of some PGRs (Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e) such as iPR and cZ in the dry extracts obtained through EAE with G1 pH4 could be explained by a more efficient and specific breakdown of seaweed cell wall polymers, thus improving the release of these compounds. This is implied by the increase in reducing sugars in G1 and G3 extracts in comparison with the WE (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e(c)). In this way, the selective depolymerization of cell wall polysaccharides and proteins tends to increase the diffusion of PGRs into the soluble phase, thus leading to higher contents for some PGRs. Moreover, two additional PGRs were quantified in the extracts (WE and EAE) in comparison with the raw seaweed. This could be explained by the temperature of extraction, the pH or the longer duration of extraction allowing a better cell wall disruption.\u003c/p\u003e \u003cp\u003eRegarding SA, the highest amount in the extract was obtained at pH 7 (WE and EAE) with significant differences observed with extracts obtained at pH 4 and pH 5. The same tendency was observed for IAA, ACC and ABA leading to the conclusion that an acidic pH has a negative effect on the recovery of these PGRs.\u003c/p\u003e \u003cp\u003eAbout cytokinins, the enzymes G1 and P2 allowed statistically significant increases in the content of iPR in comparison with extract produced by WE under the same extraction conditions. The largest increase in the content of iPR was observed in the extract obtained using the enzyme G1, which contained seven times more iPR (\u003cem\u003ep\u003c/em\u003e 0.001) compared with WE pH 4. For cZ, tZ and their precursors, in most of the extracts the values were below the lower limits of quantification of the analytical method (\u0026lt;\u0026thinsp;0.1; 0.05; 0.025 \u0026micro;g/kg d.e.), except for the enzyme G1 which enabled the production of extracts with quantified amounts of cZ, cZR and Tz. If the enzyme efficiency is compared, it appears that the extract obtained with the protease is richer in PGRs than extracts obtained with glycosidases. Indeed, P2 allowed to increase the content of iP by 8%, SA by 21%, ACC by 26%, ABA by 36%, iPR by 132% and IAA by 175% in extracts in comparison with the G3 extract.\u003c/p\u003e \u003cp\u003eResults obtained for the brown seaweed \u003cem\u003eF. vesiculosus\u003c/em\u003e in this study were in the range to those obtained by Wally et al. (\u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) for the commercial extracts produced from brown seaweeds collected in different countries, including \u003cem\u003eAscophyllum nodosum\u003c/em\u003e from France. The authors observed that the content of iP varied, ranging from 27.7 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e. for South Pacific \u003cem\u003eDurvillea\u003c/em\u003e to 1 \u0026micro;g/kg for \u003cem\u003eAscophyllum nodosum.\u003c/em\u003e It is not detected for the South African \u003cem\u003eEcklonia\u003c/em\u003e extract. cZ was detected at 1.7 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in N. Pacific \u003cem\u003eMacrocystis\u003c/em\u003e extract, while it was not detected in extracts from other countries, including \u003cem\u003eA. nodosum\u003c/em\u003e extract. The concentration of tZ was 10 \u0026micro;g/kg for Chinese \u003cem\u003eSargassum\u003c/em\u003e extract and 1 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e. for South African \u003cem\u003eEcklonia\u003c/em\u003e, with no detection in extracts from other countries, including \u003cem\u003eAscophyllum\u003c/em\u003e extract. iPR level ranged from 58.4 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for Irish soluble seaweed extract powder to non-detected for extracts from various other countries, including \u003cem\u003eAscophyllum\u003c/em\u003e extract. As for ABA, it ranged from 10 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for Chinese \u003cem\u003eSargassum\u003c/em\u003e extract to non-detected for extracts from various other countries, including French \u003cem\u003eAscophyllum\u003c/em\u003e extract. The content of IAA ranged from 615 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for Norwegian extract to 6 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e for \u003cem\u003eA. nodosum\u003c/em\u003e. Based on this study, the amount of IAA in \u003cem\u003eA. nodosum\u003c/em\u003e extracts seems to be strongly influenced by the harvesting location (country/continent) (Wally et al. \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAnother study analyzed the PGRs contents in commercial extracts produced from French Atlantic brown seaweed \u003cem\u003eA. nodosum\u003c/em\u003e (Wally et al. \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The IAA and iP contents were 6 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e. and 1 \u0026micro;g kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.e., respectively, which are much lower than the values reported in the present study. The fact remains that \u003cem\u003eA. nodosum\u003c/em\u003e has been recognized for many decades for its fertilizing and biostimulant properties.\u003c/p\u003e \u003cp\u003eThus, it appears that \u003cem\u003eF. vesiculosus\u003c/em\u003e holds promise as a raw material to develop seaweed-based product for enhancing plant growth and development. This potential stems from the diverse properties of several growth regulators found within these extracts, including their ability to improve germination, root development, and flowering, thereby supporting crucial cellular processes such as iodine uptake and energy flow within plants (Wally et al. \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Benkova 2016). Moreover, these regulators create conducive environments for plant growth, metabolism, and insect pollination (Koo et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Mattner et al. (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) conducted a study where a commercial seaweed extract derived from \u003cem\u003eDurvillaea potatorum\u003c/em\u003e and \u003cem\u003eA. nodosum\u003c/em\u003e was applied to strawberries during both nursery and production stages. Their findings demonstrated an increase in root length and density of plants during the production stage with the use of seaweed extracts. Consequently, Mattner et al. (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) suggested that seaweed extracts may enhance plant water and nutrient utilization, implying that components of seaweed other than minerals, such as PGRs, may be involved in this process. Specifically, IAA, a major PGRs present in these extracts, acts as a growth stimulant, promoting increased plant size (Benkova et al. 2016). Additionally, ABA functions as a growth inhibitor, inducing seed dormancy and influencing processes such as floral transition, lateral root formation, and seedling growth (Yang et al. 2022). Salicylic Acid, on the other hand, plays crucial roles in regulating plant disease resistance, thermogenesis, abiotic stress tolerance, DNA damage/repair, fruit yield, seed germination, and other physiological processes (Dempsey et al. 2017).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003e4.3 Evaluation of antioxidant activity with DPPH\u003c/h2\u003e \u003cp\u003eOur findings presented in Table \u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e did not reveal a straightforward relationship between antioxidant activity and TPC in \u003cem\u003eF. vesiculosus\u003c/em\u003e extracts. This result aligns with Sanchez-Bonnet et al. (2021) who described a significant negative correlation between TPC and DPPH radical scavenging activity in \u003cem\u003eF. vesiculosus\u003c/em\u003e extracts (conventional and ultrasonic assisted extraction). They found that many of their extracts containing higher and lower content of polyphenols gave lower and higher antioxidant activity, respectively (Sanchez-Bonnet et al. 2021). Similarly, Hermund et al. (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) also reported a negative correlation between TPC and RSA for \u003cem\u003eF. vesiculosus\u003c/em\u003e aqueous ethanol extracts. They proposed that the lack of a positive correlation suggests that the extraction process was not selective, and other co-extracted compounds, such as polysaccharides, pigments, proteins, peptides, or vitamins, may influence the overall antioxidant activity synergistically or antagonistically.\u003c/p\u003e \u003cp\u003eHowever, it is important to emphasize that \u003cem\u003eF\u003c/em\u003e. \u003cem\u003evesiculosus\u003c/em\u003e powder showed a total phenolic content of 39.8\u0026thinsp;\u0026plusmn;\u0026thinsp;9.8 mg PE g\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e d.s. and exhibited the lowest antioxidant activity with an IC\u003csub\u003e50\u003c/sub\u003e of 309.2 \u0026micro;g mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e compared to the different extraction techniques applied in this study. Since the total phenolic content found with P2 enzyme at pH 7 was significantly greater than the content in the seaweed powder, this suggests that EAE with P2 not only enhance the overall polyphenol content in the extracts, but also improve the antioxidant activity in \u003cem\u003eF. vesiculosus\u003c/em\u003e by lowering their IC\u003csub\u003e50\u003c/sub\u003e value.\u003c/p\u003e \u003c/div\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eThis study aimed to develop and evaluate the effectiveness of two extraction methods \u0026mdash; Enzyme Assisted Extraction (EAE) and Water Extraction (WE) \u0026mdash; for the valorization of \u003cem\u003eFucus vesiculosus\u003c/em\u003e Linnaeus. The findings indicated that pH influenced the recovery of compounds from \u003cem\u003eF. vesiculosus\u003c/em\u003e, with EAE showing higher efficiency at pH 4 compared to pH 5 and pH 7. Although EAE at pH 4 had a higher extraction yield than WE at the same pH, the difference was not significant. Similarly, WE was more efficient at pH 4 than at pH 5 and pH 7. Furthermore, this study demonstrated that EAE was significantly more effective than WE in producing enriched extracts, enhancing the release of bioactive compounds such as total sugars, a few plant growth regulators (PGRs), and polyphenols. Additionally, the polyphenol content and antioxidant activity of the raw algal powder and the various extracts were compared. The results highlighted that the extracts exhibited superior antioxidant activity, which can play a crucial role in agriculture by enhancing plant health through protection against oxidative stress, thereby supporting sustainable and eco-friendly agricultural practices by reducing reliance on chemical inputs.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThis research was supported by the National Science Centre in Poland [grant 2019/33/B/NZ9/01844]; the French government under the programme \u0026ldquo;Investissements d\u0026apos;Avenir\u0026rdquo; - France 2030; Agro Innovation International \u0026ndash; Timac Agro; and the ISblue project [grant ANR- 17-EURE-0015]. The authors thank Damien Souquet for the LC-MS analyses of PGRs.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval\u003c/strong\u003e Not applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003eThis research was supported by the National Science Centre in Poland [grant 2019/33/B/NZ9/01844]; the French government under the programme \u0026ldquo;Investissements d\u0026apos;Avenir\u0026rdquo; - France 2030; Agro Innovation International \u0026ndash; Timac Agro; and the ISblue project [grant ANR- 17-EURE-0015].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u0026nbsp;\u003c/strong\u003eThe data that support the findings of this study are available from the corresponding author, (Nathalie Bourgougnon).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of competing interest\u003c/strong\u003e The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAdams, JMM, Ross AB, Anastasakis K, Hodgson E.M, Gallagher JA, Jones JM, Donnison IS (2011) Seasonal variation in the chemical composition of the bioenergy feedstock \u003cem\u003eLaminaria digitata\u003c/em\u003e for thermochemical conversion. Bioresource Technol, Special Issue: Biofuels - II: Algal Biofuels and Microbial Fuel Cells 102:226\u0026ndash;234. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.biortech.2010.06.152\u003c/span\u003e\u003cspan address=\"10.1016/j.biortech.2010.06.152\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAgreg\u0026aacute;n R, Munekata PE, Dom\u0026iacute;nguez R, Carballo J, Franco D, Lorenzo JM (2017) Proximate composition, phenolic content and in vitro antioxidant activity of aqueous extracts of the seaweeds Ascophyllum nodosum, Bifurcaria bifurcata and Fucus vesiculosus. Effect of addition of the extracts on the oxidative stability of canola oil under accelerated storage conditions. Food Res Int 99:986\u0026ndash;994.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.foodres.2016.11.009\u003c/span\u003e\u003cspan address=\"10.1016/j.foodres.2016.11.009\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAllen E, Wall DM, Herrmann C, Xia A, Murphy JD (2015) What is the gross energy yield of third generation gaseous biofuel sourced from seaweed? Energy 81:352\u0026ndash;360. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.energy.2014.12.048\u003c/span\u003e\u003cspan address=\"10.1016/j.energy.2014.12.048\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArzel P (1987) Les Go\u0026eacute;moniers. Le Chasse-Mar\u0026eacute;e, Douarnenez. ISBN 2903708053\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBalina K, Romagnoli F, Blumberga D (2016) Chemical Composition and Potential Use of Fucus vesiculosus from Gulf of Riga. Energy Proced 95:43\u0026ndash;49.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.egypro.2016.09.010\u003c/span\u003e\u003cspan address=\"10.1016/j.egypro.2016.09.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBenkov\u0026aacute; E (2016) Plant hormones in interactions with the environment. Plant Mol Biol 91:597.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11103-016-0501-8\u003c/span\u003e\u003cspan address=\"10.1007/s11103-016-0501-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBenitez Garc\u0026iacute;a I, Due\u0026ntilde;as Ledezma AK, Mart\u0026iacute;nez-Monta\u0026ntilde;o E., Salazar Leyva JA, Carrera E, Osuna Ruiz I (2020) Identification and Quantification of Plant Growth Regulators and Antioxidant Compounds in Aqueous Extracts of Padina durvillaei and Ulva lactuca. Agronomy, 10: 866. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agronomy10060866\u003c/span\u003e\u003cspan address=\"10.3390/agronomy10060866\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBesednova NN, Andryukov BG, Zaporozhets TS, Kryzhanovsky SP, Kuznetsova TA, Fedyanina LN, Makarenkova ID, Zvyagintseva TN (2020) Algae polyphenolic compounds and modern antibacterial strategies: Current achievements and immediate prospects. Biomedicines 8:342. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/biomedicines8090342\u003c/span\u003e\u003cspan address=\"10.3390/biomedicines8090342\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBlumenkrantz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem. 54(2):484\u0026ndash;9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0003-2697(73)90377-1\u003c/span\u003e\u003cspan address=\"10.1016/0003-2697(73)90377-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBikovens O, Ponomarenko J, Janceva S, Lauberts M, Vevere L, Telysheva G (2017) Development of the Approaches for Complex Utilization of brown algae (\u003cem\u003eFucus vesiculosus\u003c/em\u003e) Biomass for the Obtaining of Value-Added Products. Proceedings of the 8th International Scientific Conference Rural Development. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.15544/RD.2017.091\u003c/span\u003e\u003cspan address=\"10.15544/RD.2017.091\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBuggeln RG, Craigie JS (1971) Evaluation of evidence for the presence of indole-3-acetic acid in marine algae. Planta 97:173\u0026ndash;178. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF00386764\u003c/span\u003e\u003cspan address=\"10.1007/BF00386764\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBunker F, Brodie JA, Maggs CA, Bunker AR (2017) Seaweeds of Britain and Ireland: Second Edition. Wild nature press. ISBN 9780995567337\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCatarino MD, Silva AMS, Cardoso SM (2017) Review Fucaceae: A Source of Bioactive Phlorotannins. Int J Mol Sci 18:1327. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ijms18061327\u003c/span\u003e\u003cspan address=\"10.3390/ijms18061327\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCatarino, MD, Silva, AMS, Cardoso SM (2018) Phycochemical constituents and biological activities of \u003cem\u003eFucus\u003c/em\u003e spp. Mar Drugs 16:249. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/md16080249\u003c/span\u003e\u003cspan address=\"10.3390/md16080249\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCatarino MD, Silva AMS, Mateus N, Cardoso SM (2019) Optimization of phlorotannins extraction from \u003cem\u003eFucus vesiculosus\u003c/em\u003e and evaluation of their potential to prevent metabolic disorders. Mar Drugs 17(3):162. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/md17030162\u003c/span\u003e\u003cspan address=\"10.3390/md17030162\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChambers JM, Hastie TJ (1992) Statistical Models in S, Wadsworth \u0026amp; Brooks/Cole\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoulot M, Le Guillard C, Bourgougnon N, Michalak I (2022) Seaweeds-based fertilizing products in: \u0026ldquo;Algae and Aquatic Macrophytes in Cities\u0026rdquo;. Vimal Chandra Pandey, ISBN: 978-0-12-824270-4.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoulot M, Michalak I, Jing L, Szymczycha-Madeja A, Wełna M, Bourgougnon N, Le Guillard C (2023) The Enzyme-Assisted Extraction of compounds of interest in agriculture: Case study of the red seaweed \u003cem\u003eSolieria chordalis\u003c/em\u003e (C. Agardh) J. Agardh, Algal Research 75:103239.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.algal.2023.103239\u003c/span\u003e\u003cspan address=\"10.1016/j.algal.2023.103239\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDempsey DA, Klessig DF (2017) How does the multifaceted plant hormone salicylic acid combat disease in plants and are similar mechanisms utilized in humans? BMC Biol 15:23.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s12915-017-0364-8\u003c/span\u003e\u003cspan address=\"10.1186/s12915-017-0364-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDeniaud-Bou\u0026euml;t E, Hardouin K, Potin P, Kloareg B, Herv\u0026eacute;, C (2017) A review about brown algal cell walls and fucose-containing sulfated polysaccharides: Cell wall context, biomedical properties and key research challenges. Carbohyd Polym 175:395\u0026ndash;408.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.carbpol.2017.07.082\u003c/span\u003e\u003cspan address=\"10.1016/j.carbpol.2017.07.082\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eD\u0026iacute;az-Rubio ME, P\u0026eacute;rez-Jim\u0026eacute;nez J, Saura-Calixto F (2009) Dietary fiber and antioxidant capacity in \u003cem\u003eFucus vesiculosus\u003c/em\u003e products. Int J Food Sci. Nutr 60:23\u0026ndash;34. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/09637480802189643\u003c/span\u003e\u003cspan address=\"10.1080/09637480802189643\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDuBois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric Method for Determination of Sugars and Related Substances. Anal Chem 28:350\u0026ndash;356. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/ac60111a017\u003c/span\u003e\u003cspan address=\"10.1021/ac60111a017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFilisetti-Cozzi TMCC, Carpita NC (1991) Measurement of uronic acids without interference from neutral sugars. Anal Biochem 197: 157\u0026ndash;162. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0003-2697(91)90372-Z\u003c/span\u003e\u003cspan address=\"10.1016/0003-2697(91)90372-Z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFleurence J (1999) Seaweed proteins: biochemical, nutritional aspects and potential uses. Trends Food Sci Tech 10:25\u0026ndash;28. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0924-2244(99)00015-1\u003c/span\u003e\u003cspan address=\"10.1016/S0924-2244(99)00015-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFox J, Bouchet-Valat M, Marquez MM, et al. (2024) Rcmdr: R Commander. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://cran.r-project.org/web/packages/Rcmdr/index.html\u003c/span\u003e\u003cspan address=\"https://cran.r-project.org/web/packages/Rcmdr/index.html\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e; accessed 8 February 2024\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGall EA, Lelchat F, Hupel M, J\u0026eacute;gou C, Stiger-Pouvreau V (2015) Extraction and Purification of Phlorotannins from Brown Algae. In: Stengel, D., Connan, S. (eds) Natural Products From Marine Algae. Methods in Molecular Biology, vol 1308. Humana Press, New York. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/978-1-4939-2684-8_7\u003c/span\u003e\u003cspan address=\"10.1007/978-1-4939-2684-8_7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHaddad C, Arkoun M, Jamois F, Schwarzenberg A, Yvin J-C, Etienne P and La\u0026icirc;n\u0026eacute; P (2018) Silicon promotes growth of \u003cem\u003eBrassica napus\u003c/em\u003e L. and delays leaf senescence induced by nitrogen starvation. Front Plant Sci. 9:516. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2018.00516\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2018.00516\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHagerman AE, Butler LG (1989) Choosing appropriate methods and standards for assaying tannin. J Chem Ecol 15:1795\u0026ndash;1810. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01012267\u003c/span\u003e\u003cspan address=\"10.1007/BF01012267\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHardouin K, Bedoux G, Burlot AS, Nyvall-Collen P, Bourgougnon N (2014) Enzymatic recovery of metabolites from seaweeds: potential applications. Adv Bot Res 71:279\u0026ndash;320. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/B978-0-12-408062-1.00010-X\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-12-408062-1.00010-X\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHardouin K, Burlot AS, Umami A, Tanniou A, Stiger-Pouvreau V, Widowati I, Bedoux G, Bourgougnon N (2014) Biochemical and antiviral activities of enzymatic hydrolysates from different invasive French seaweeds. J Appl Phycol 26:1029\u0026ndash;1042. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10811-013-0201-6\u003c/span\u003e\u003cspan address=\"10.1007/s10811-013-0201-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHermund DB, Plaza M, Turner C, J\u0026oacute;nsd\u0026oacute;ttir R, Kristinsson HG, Jacobsen C, Jacobsen C, Fog Nielsen K (2018) Structure dependent antioxidant capacity of phlorotannins from Icelandic \u003cem\u003eFucus vesiculosus\u003c/em\u003e by UHPLC-DAD-ECD-QTOFMS. Food Chem 240:904\u0026ndash;909. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.foodchem.2017.08.032\u003c/span\u003e\u003cspan address=\"10.1016/j.foodchem.2017.08.032\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHollander M, Wolfe DA, Chicken E (1973). Nonparametric Statistical Methods. Wiley, New York\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJacob C, Frangoudes K, Mesnildrey L, Lesueur M, Spinec F, Folliard G, Pien S, Rebours C, Meland M, Gallastegi I, Inunziaga Urizarbarrena A, Marques A, Atack T, Dunningham J, Maguire J, Abreu H, Walsh M (2012) Etats des lieux de la fili\u0026egrave;re des macro-algues en Europe. projet NETALGAE - Interreg IVB.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWorld Bank (2023) Global Seaweed: New and Emerging Markets Report, 2023. \u0026copy; Washington, DC:World Bank. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://hdl.handle.net/10986/40187\u003c/span\u003e\u003cspan address=\"http://hdl.handle.net/10986/40187\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKadam SU, Tiwari BK, O\u0026rsquo;Donnell CP (2013) Application of novel extraction technologies for bioactives from marine algae. J. Agric. Food Chem. 61:4667\u0026ndash;4675. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/jf400819p\u003c/span\u003e\u003cspan address=\"10.1021/jf400819p\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKnight M, Parke M (1950) A biological study of \u003cem\u003eFucus vesiculosus\u003c/em\u003e L. and \u003cem\u003eFucus serratus L\u003c/em\u003e. J Mar Biol Assoc UK 29:439\u0026ndash;514. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0025315400055454\u003c/span\u003e\u003cspan address=\"10.1017/S0025315400055454\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoo YM, Heo AY, Choi HW (2020) Salicylic Acid as a Safe Plant Protector and Growth Regulator. Plant Pathol J 36(1):1\u0026ndash;10. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5423/PPJ.RW.12.2019.0295\u003c/span\u003e\u003cspan address=\"10.5423/PPJ.RW.12.2019.0295\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKrautforst K, Szymczycha-Madeja A, Welma M, Michalak I (2023) Brown seaweed: \u003cem\u003eFucus vesiculosus\u003c/em\u003e as a feedstock for agriculture and environment protection. Sci Rep 13, 100065 \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41598-023-36881-z\u003c/span\u003e\u003cspan address=\"10.1038/s41598-023-36881-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLaekeman, G (2014) Assessment Report on \u003cem\u003eFucus vesiculosus\u003c/em\u003e L., Thallus \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ema.europa.eu/en/documents/herbal-report/final-assessment-report-fucus-vesiculosus-l-thallus_en.pdf\u003c/span\u003e\u003cspan address=\"https://www.ema.europa.eu/en/documents/herbal-report/final-assessment-report-fucus-vesiculosus-l-thallus_en.pdf\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLe Guillard C, Berg\u0026eacute;, JP, Donnay-Moreno C, Cornet J, Ragon JY, Fleurence J, Dumay J (2023) Optimization of R-Phycoerythrin Extraction by Ultrasound-Assisted Enzymatic Hydrolysis: A Comprehensive Study on the Wet Seaweed \u003cem\u003eGrateloupia turuturu\u003c/em\u003e. Mar Drugs 21:213. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/md21040213\u003c/span\u003e\u003cspan address=\"10.3390/md21040213\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLe Lann K, J\u0026eacute;gou C, Stiger-Pouvreau V (2008) Effect of different conditioning treatments on total phenolic content and antioxidant activities in two Sargassacean species: Comparison of the frondose Sargassum muticum (Yendo) Fensholt and the cylindrical Bifurcaria bifurcata R. Ross. Phycol Res 56: 238\u0026ndash;245. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1440-1835.2008.00505.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1440-1835.2008.00505.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi J, Chen K, Li S, Liu T, Wang F, Xia Y, Lu J, Zhou Y, Guo C (2016) Pretreatment with Fucoidan from \u003cem\u003eFucus vesiculosus\u003c/em\u003e Protected against ConA-Induced Acute Liver Injury by Inhibiting Both Intrinsic and Extrinsic Apoptosis. PLOS One 11(4):e0152570. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0152570\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0152570\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMaheswaria V, Babu PA S (2022) Phlorotannin and its derivatives, a potential antiviral molecule from brown seaweeds, an overview. Russ J Mar Biol\u0026thinsp;+\u0026thinsp;48 (5):309\u0026ndash;324. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1134/S1063074022050169\u003c/span\u003e\u003cspan address=\"10.1134/S1063074022050169\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMahmoud E, Abd El-Kader N (2015) Heavy metal immobilization in contaminated soils using phosphogypsum and rice straw compost. Land Degrad Dev 26:819\u0026ndash;824. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/ldr.2288\u003c/span\u003e\u003cspan address=\"10.1002/ldr.2288\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMattner SW, Milinkovic M, Arioli T (2018) Increased growth response of strawberry roots to a commercial extract from \u003cem\u003eDurvillaea potatorum\u003c/em\u003e and \u003cem\u003eAscophyllum nodosum\u003c/em\u003e. J Appl Phycol 30:2943\u0026ndash;2951.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10811-017-1387-9\u003c/span\u003e\u003cspan address=\"10.1007/s10811-017-1387-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMesnildrey L, Jacob C\u0026eacute;line, Frangoudes K, Reunavot M, Lesueur M (2012) Seaweed industry in France. Report Interreg program NETALGAE. 2012, 42 p. ⟨hal-00840572⟩\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMichalak I, Chojnacka K (2014) Algal extracts: Technology and advances. Eng Life Sci 14: 581\u0026ndash;591. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/elsc.201400139\u003c/span\u003e\u003cspan address=\"10.1002/elsc.201400139\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMichalak I, Baśladyńska S (2021) Effect of Fucus extract and biomass enriched with Cu (II) and Zn (II) ions on the growth of garden cress (Lepidium sativum) under laboratory conditions. Ital J Agron 16: 1799. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4081/ija.2021.1799\u003c/span\u003e\u003cspan address=\"10.4081/ija.2021.1799\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31(3):426\u0026ndash;428. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1021/ac60147a030\u003c/span\u003e\u003cspan address=\"10.1021/ac60147a030\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMu X, Chen Y (2021) The physiological response of photosynthesis to nitrogen deficiency. Plant Physiol Bioch 158:76\u0026ndash;82. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.plaphy.2020.11.019\u003c/span\u003e\u003cspan address=\"10.1016/j.plaphy.2020.11.019\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMzibra, A., Aasfar, A., Benhima, R. et al. Biostimulants derived from Moroccan seaweeds: seed germination metabolomics and growth promotion of tomato plant. J Plant Growth Regul 40, 353\u0026ndash;370 (2021). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00344-020-10104-5\u003c/span\u003e\u003cspan address=\"10.1007/s00344-020-10104-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNeiva J, Serr\u0026atilde;o EA, Assis J, Pearson GA, Coyer JA, Olsen JL, Hoarau G, Valero M (2016) Climate oscillations, range shifts and phylogeographic patterns of North Atlantic Fucaceae. In: Hu Z-M, Fraser C (eds) Seaweed Phylogeography. Springer Netherlands, Dordrecht, pp279\u0026ndash;308\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNeto RT, Mar\u0026ccedil;al C, Queir\u0026oacute;s AS, Abreu H, Silva AMS, Cardoso SM (2018) Screening of \u003cem\u003eUlva rigida\u003c/em\u003e, \u003cem\u003eGracilaria\u003c/em\u003e sp., \u003cem\u003eFucus vesiculosus\u003c/em\u003e and \u003cem\u003eSaccharina latissima\u003c/em\u003e as Functional Ingredients. Int J Mol Sci 19:2987. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ijms19102987\u003c/span\u003e\u003cspan address=\"10.3390/ijms19102987\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eObluchinskaya ED (2008) Comparative chemical composition of the Barents Sea brown algae. Appl Biochem Microbiol 44:305\u0026ndash;309. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1134/S0003683808030149\u003c/span\u003e\u003cspan address=\"10.1134/S0003683808030149\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eObluchinskaya ED, Pozharitskaya ON, Zakharov DV, Flisyuk EV, Terninko II, Generalova YE, Smekhova IE, Shikov AN (2022) The Biochemical Composition and Antioxidant Properties of \u003cem\u003eFucus vesiculosus\u003c/em\u003e from the Arctic Region. Mar Drugs 20:193. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/md20030193\u003c/span\u003e\u003cspan address=\"10.3390/md20030193\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOsoski AL, Lohr P, Reiff M, Balick MJ, Kronenberg F, Fugh-Berman A, O\u0026rsquo;Connor B (2002) Ethnobotanical literature survey of medicinal plants in the Dominican Republic used for women\u0026rsquo;s health conditions. J Ethnopharmacol 79:285\u0026ndash;298.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003e\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0378-8741(01)00376-2\u003c/span\u003e\u003cspan address=\"10.1016/S0378-8741(01)00376-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOverbeek, JV (1940) Auxin in Marine Algae. Plant Physiol 15:291\u0026ndash;299. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1104/pp.15.2.291\u003c/span\u003e\u003cspan address=\"10.1104/pp.15.2.291\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePaiva L., Lima E., Neto A.I., Baptista J. Seasonal variability of the biochemical composition and antioxidant properties of Fucus spiralis at two Azorean islands. Mar. Drugs 2018, 16, 248; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/md16080248\u003c/span\u003e\u003cspan address=\"10.3390/md16080248\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eParys S, Kehraus S, Krick A, Glombitza KW, Carmeli S, Klimo K, K\u0026ouml;nig GM (2010) \u003cem\u003eIn vitro\u003c/em\u003e chemopreventive potential of fucophlorethols from the brown alga \u003cem\u003eFucus vesiculosus L.\u003c/em\u003e by anti-oxidant activity and inhibition of selected cytochrome P450 enzymes. Phytochemistry 71:221\u0026ndash;229. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.phytochem.2009.10.020\u003c/span\u003e\u003cspan address=\"10.1016/j.phytochem.2009.10.020\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePierre G (2010). Biochemical characterization of algal exopolymers from Marennes-Ol\u0026eacute;ron and study of the physico-chemical surface properties of microorganisms involved in their adhesion (phdthesis). Universit\u0026eacute; de La Rochelle.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR\u0026eacute;thor\u0026eacute; E, Jing L, Ali N, Yvin JC, Pluchon S, Hosseini SA (2021) K Deprivation modulates the primary metabolites and increases putrescine concentration in \u003cem\u003eBrassica napus\u003c/em\u003e. Front Plant Sci 12:681895. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2021.681895\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2021.681895\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRodr\u0026iacute;guez-Jasso RM, Mussatto SI, Pastrana L, Aguila CN, Teixeira JA (2013) Extraction of sulfated polysaccharides by autohydrolysis of brown seaweed \u003cem\u003eFucus vesiculosus\u003c/em\u003e. J Appl Phycol 25:31\u0026ndash;39. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10811-012-9834-0\u003c/span\u003e\u003cspan address=\"10.1007/s10811-012-9834-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRomm A, Hardy ML, Mills S, Bove M, Stansbury JE, Romm A (2010) Chap. 6 Endocrine Disorders and Adrenal Support. In: Botanical Medicine for Women\u0026rsquo;s Health; Churchill Livingstone: London, UK, pp 186\u0026ndash;210. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/B978-0-443-07277-2.X0001-3\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-443-07277-2.X0001-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRousseau C, Demoulinger G, Rousvoal S, Champeval D, Dolly M, Michel G, Thomas F, Markov G.V, Potin P, Dittami S.M, Prado S, Leblanc C, Le Guillard C (2023) A review on the chemical ecology of the Fucaceae holobionts: from fundamental knowledge to applications. CR Chim 26:1\u0026ndash;25. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5802/crchim.271\u003c/span\u003e\u003cspan address=\"10.5802/crchim.271\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaebmehr H, Rafiee F, Givianrad M H, Mostafavi G (2022) Determination and quantification of two regulatory phytohormones extracted from \u003cem\u003eSargassum muticum\u003c/em\u003e and \u003cem\u003eGracilaria corticate\u003c/em\u003e seaweeds in the south of Iran. Nova Biologica Reperta 9(2):115\u0026ndash;123. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.29252/nbr.9.2.115\u003c/span\u003e\u003cspan address=\"10.29252/nbr.9.2.115\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSanchez-Bonet D, Garc\u0026iacute;a-Oms S, Belda-Antol\u0026iacute; M, Padr\u0026oacute;n-Sanz C, Lloris-Carsi JM, Cejalvo-Lape\u0026ntilde;a D (2021) RP-HPLC-DAD determination of the differences in the polyphenol content of Fucus vesiculosus extracts with similar antioxidant activity. J Chromatogr B 1184:122978. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jchromb.2021.122978\u003c/span\u003e\u003cspan address=\"10.1016/j.jchromb.2021.122978\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma HSS, Lyons G, McRoberts C, McCall D, Carmichael E, Andrews F, Swan R (2012a) Biostimulant activity of brown seaweed species from Strangford Lough: compositional analyses of polysaccharides and bioassay of extracts using mung bean (Vigno mungo L.) and pak choi (Brassica rapa chinensis L.). J Appl Phycol 24:1081\u0026ndash;1091. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10811-011-9737-5\u003c/span\u003e\u003cspan address=\"10.1007/s10811-011-9737-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSharma HSS, Fleming C, Selby C, Rao JR, Martin T (2014) Plant biostimulants: a review on the processing of macroalgae and use of extracts for crop management to reduce abiotic and biotic stresses. J Appl Phycol 26:465\u0026ndash;490. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10811-013-0101-9\u003c/span\u003e\u003cspan address=\"10.1007/s10811-013-0101-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingleton VL, Orthofer R, Lamuela-Raventos RM (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu Reagent. Method Enzymol 299: 152\u0026ndash;178. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0076-6879(99)99017-1\u003c/span\u003e\u003cspan address=\"10.1016/S0076-6879(99)99017-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSiriwardhana N, Kim KN, Lee KW, Kim SH, Ha JH, Song CB, Lee JB, Jeon YJ (2008) Optimisation of hydrophilic antioxidant extraction from Hizikia fusiformis by integrating treatments of enzymes, heat and pH control. Int J Food Sci Tech 43(4):587\u0026ndash;596. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1365-2621.2006.01485.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1365-2621.2006.01485.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSkibola CF (2004) The effect of \u003cem\u003eFucus vesiculosus\u003c/em\u003e, an edible brown seaweed, upon menstrual cycle length and hormonal status in three pre-menopausal women: A case report. BMC Complement. Altern Med 4:10. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1472-6882-4-10\u003c/span\u003e\u003cspan address=\"10.1186/1472-6882-4-10\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSmith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem, 150(1):76\u0026ndash;85.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0003-2697(85)90442-7\u003c/span\u003e\u003cspan address=\"10.1016/0003-2697(85)90442-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSpain O, Hardouin K, Bourgougnon N, Michalak I (2024) Enzyme Assisted Extraction of red seaweed \u003cem\u003eSolieria chordalis\u003c/em\u003e) J. Agardh 184 - the starting point for the production of biostimulants of plant growth and biosorbents of metal ions. Biomass Conversion and Biorefinery Journal 14:1621\u0026ndash;1635. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s13399-022-02456-7\u003c/span\u003e\u003cspan address=\"10.1007/s13399-022-02456-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShukla PS, Borza T, Critchley AT, Prithiviraj B (2021) Seaweed-Based Compounds and Products for Sustainable Protection against Plant Pathogens. Mar Drugs 19(2):59. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/md19020059\u003c/span\u003e\u003cspan address=\"10.3390/md19020059\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStirk W, Novak O, Strnad M, Staden J (2003) Cytokinins in macroalgae. Plant Growth Regul 41:13\u0026ndash;24. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1023/A:1027376507197\u003c/span\u003e\u003cspan address=\"10.1023/A:1027376507197\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStirk WA, Van Staden J (2014) Chapter Five: Plant Growth Regulators in Seaweeds: Occurrence, Regulation and Functions. In: Bourgougnon, N. (Ed.), Advances in Botanical Research, Sea Plants. Academic Press, pp 125\u0026ndash;159. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/B978-0-12-408062-1.00005-6\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-12-408062-1.00005-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTeixeira-Guedes C, Gomes-Dias JS, Cunha SA, Pintado ME, Pereira RN, Teixeira JA, Rocha CMR (2023) Enzymatic approach for the extraction of bioactive fractions from red, green and brown seaweeds. Food Bioprod Process 138:25\u0026ndash;39. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fbp.2022.12.005\u003c/span\u003e\u003cspan address=\"10.1016/j.fbp.2022.12.005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWally OSD, Critchley AT, Hiltz D, Craigie JS, Han X, Zaharia LI, Abrams SR, Prithiviraj B (2012) Regulation of Phytohormone Biosynthesis and Accumulation in Arabidopsis Following Treatment with Commercial Extract from the Marine Macroalga Ascophyllum nodosum. J Plant Growth Regul 32:324\u0026ndash;339. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00344-012-9301-9\u003c/span\u003e\u003cspan address=\"10.1007/s00344-012-9301-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang M, Zheng Q, Shen Q, Guo S (2013) The Critical Role of Potassium in Plant Stress Response. Int J Mol Sci 14(4): 7370\u0026ndash;7390. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/ijms14047370\u003c/span\u003e\u003cspan address=\"10.3390/ijms14047370\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang T, J\u0026oacute;nsd\u0026oacute;ttir R, Kristinsson HG, Hreggvidsson GO, J\u0026oacute;nsson J\u0026Oacute;, Thorkelsson G, \u0026Oacute;lafsd\u0026oacute;ttir G (2010) Enzyme-enhanced extraction of antioxidant ingredients from red algae Palmaria palmata. LWT - Food Sci Technol 43:1387\u0026ndash;1393. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.lwt.2010.05.010\u003c/span\u003e\u003cspan address=\"10.1016/j.lwt.2010.05.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWaterhouse AL (2002) Determination of Total Phenolics. Current Protocols in Food Analytical Chemistry, I1.1.1-I\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e1.1.8\u003c/span\u003e\u003cspan address=\"http://1.1.8\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/0471142913.fai0101s06\u003c/span\u003e\u003cspan address=\"10.1002/0471142913.fai0101s06\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWijesinghe WAJP, Jeon YJ (2012) Enzyme-assistant extraction (EAE) of bioactive components: A useful approach for recovery of industrially important metabolites from seaweeds: A review. Fitoterapia 83:6\u0026ndash;12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fitote.2011.10.016\u003c/span\u003e\u003cspan address=\"10.1016/j.fitote.2011.10.016\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWissuwa M (2003) How Do Plants Achieve Tolerance to Phosphorus Deficiency? Small Causes with Big Effects. Plant Physiol 133(4):1947\u0026ndash;1958. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1104/pp.103.029306\u003c/span\u003e\u003cspan address=\"10.1104/pp.103.029306\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYalcin S, Okudan E, Karakaş \u0026Ouml;, \u0026Ouml;nem A, S\u0026ouml;zgen Başkan K (2019) Identification and quantification of some phytohormones in seaweeds using UPLC-MS/MS. J Liq Chromatogr R T 42:1\u0026ndash;10. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1080/10826076.2019.1625374\u003c/span\u003e\u003cspan address=\"10.1080/10826076.2019.1625374\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"","identity":"journal-of-applied-phycology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"10811","submissionUrl":"https://submission.nature.com/new-submission/10811/3","title":"Journal of Applied Phycology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Brown seaweed, Fucus vesiculosus, Enzyme-Assisted Extraction, Biochemical Composition, Elements, Plant Growth Regulators","lastPublishedDoi":"10.21203/rs.3.rs-5331195/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5331195/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe agricultural sector plays a crucial role in sustaining population growth and ensuring our well-being. However, as this sector faces numerous challenges due to environmental conditions and the increasing population, it is essential to identify natural alternatives that promote sustainable farming practices and protect the environment. Seaweed-based extracts have gained popularity in agriculture because of their numerous benefits for plant growth and health. This study focuses on \u003cem\u003eFucus vesiculosus\u003c/em\u003e Linnaeus, a prevalent brown seaweed species found along the Brittany coast. The research involved comparing Enzyme-Assisted Extraction (EAE) with conventional aqueous extraction methods for the purpose of determining the effectiveness of EAE in producing enriched extracts that could have an interesting agricultural application. The results indicates that enzymatic extraction of \u003cem\u003eFucus vesiculosus\u003c/em\u003e significantly increased the content of neutral sugars by 34% and reducing sugars by 21% in the extracts, compared to conventional aqueous extraction (WE). Regarding Plant Growth Regulators, the levels of Isopentenyladenosine (iPR) and Cis zeatin (cZ) were enhanced by 6 times and 28 times, respectively, when using EAE instead of WE. Additionally, the total phenolic content was notably higher in EAE extracts, showing a twofold increase over WE extracts. Furthermore, the various extracts demonstrated superior antioxidant activity compared to raw \u003cem\u003eFucus vesiculosus\u003c/em\u003e powder. Thus, this study confirms that EAE is an effective method for enriching \u003cem\u003eFucus vesiculosus\u003c/em\u003e extracts with various compounds that can play a vital role in agriculture.\u003c/p\u003e","manuscriptTitle":"Application of Enzyme-Assisted Extraction on the brown seaweed Fucus vesiculosus Linnaeus (Ochrophyta, Fucaceae) to produce extracts for a sustainable agriculture.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-07 12:02:48","doi":"10.21203/rs.3.rs-5331195/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-12-12T05:47:38+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-12-12T02:33:38+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-11-25T17:10:53+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"336853303643006942011642418549918412706","date":"2024-11-07T02:22:21+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"323878199457232327198923812923347593164","date":"2024-11-04T16:24:44+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-11-03T09:40:33+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-10-29T01:27:19+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-10-29T00:42:49+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Applied Phycology","date":"2024-10-25T09:19:59+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"","identity":"journal-of-applied-phycology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"10811","submissionUrl":"https://submission.nature.com/new-submission/10811/3","title":"Journal of Applied Phycology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"371b4460-013d-40ff-9d92-457763997a45","owner":[],"postedDate":"November 7th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2024-12-27T07:38:38+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-07 12:02:48","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5331195","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5331195","identity":"rs-5331195","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}