Grewia asiatica leaves extract assisted green synthesis of gold nanoparticles and study of their antibacterial, antioxidant and photocatalytic potential

Grewia asiatica leaves extract assisted green synthesis of gold nanoparticles and study of their antibacterial, antioxidant and photocatalytic potential | 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 Grewia asiatica leaves extract assisted green synthesis of gold nanoparticles and study of their antibacterial, antioxidant and photocatalytic potential Sana Maryam, Naseem Akhter, Muhammad Azam Qamar, Asma Yaqoob, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3798283/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The environmental issues generated by industrial advancement and human activities are serious and must not be overlooked. Bio-nanotechnology has emerged as a solution for producing biosynthetic nanomaterials to address environmental issues. The current work describes for the first time an economical and environmentally benign method of producing gold nanoparticles utilizing Grewia asiatica (GAAuNPs). The optimal AuNPs were produced using a gold ion concentration of 1 mM and a metal-to-extract ratio of 1:1. The resulting nanoparticles were analyzed and characterized using X-ray crystallography, FTIR, SEM, EDX, and UV visible spectroscopy. The catalytic methylene blue (MB) dye degradation properties, antibacterial and biofilm inhibition abilities, and antioxidant properties of the synthesized particles were investigated. The Au NPs demonstrated efficient catalytic degradation against MB and completely degraded it in 8 minutes. The NPs also exhibited potent biofilm inhibition against E. coli and S. aureus that was close to the standard and was more antioxidant than the standard. The findings highlight G. asiatica as a suitable, inexpensive biosource for biofabricating GAAuNPs with extensive multifunctional uses. To conclude, GAAuNPs' ability to degrade organic pollutants and eradicate pathogens provides a cost-effective and environmentally benign remedy to tackle contemporary pollutants. Green nanoparticles dye degradation biofilm inhibition antibacterial activity Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 1. Introduction Due to the global threat of resistant bacteria, food safety, economic growth, and human and animal health are all at risk (Man et al. 2019 ; Puvača et al. 2021 ). Additionally, bacteria significantly contribute to persistent and recurring infections because of their propensity to create biofilms, which shield them from the host's immune system, antibiotics, and disinfectants (Singh et al. 2018 ; Nassima et al. 2019 ). In today's world, it is essential to have an efficient and advanced plan for managing and treating drug-resistant microorganisms (Swolana et al. 2020 ). Similarly, an unfavorable ecological shift is accompanied by untreated industrial and domestic effluents being discharged into the ecosystem(Natasha et al. 2020). The waste dye molecules significantly negatively impact humans, wildlife, and vegetation. As a result, organic dye effluents are now recognized as a significant threat to aquatic environments (Iqbal et al. 2020 ; Ahmad et al. 2022 ). Organic dyes are the most hazardous of all the contaminants released into our environment. The primary contaminants in industrial effluent are dyes because of their color (Javed et al. 2021 ; Sher et al. 2021a ). The biological breakdown of dye molecules is exceedingly difficult because they are tenacious compounds. Due to their toxicity, commercial significance, and environmental effects, dyes have been the subject of extensive research (Qamar et al. 2022b ). As a result, pathogens and organic contaminants must be removed and detoxified from water bodies more urgently than other pollutants. Different remediation techniques, including chemical, physiochemical, and biological treatments, have so far been suggested. However, each has its pros and cons compared to the others(Sher et al. 2021a ; Javed et al. 2023 ). These colored pollutants are difficult to remove using traditional methods (such as adsorption, coagulation, flocculation, biodegradation, and so on), and all of these procedures are expensive and require additional planning to remove the byproducts. Alternatively, photocatalytic degradation of hazardous dyes into harmless chemicals is the preferred method for reducing their environmental impact. Developing an effective and long-term solution for disposing of harmful dyes and pathogens in wastewater streams is paramount. Recent advances in nanotechnology have enabled the use of nanoparticles as biological agents for suppressing bacteria and as catalysts for degrading toxic pollutants. Materials at the nanoscale are the focus of nanoscience, which has applications in many different areas, including medicine, engineering, forensics, agriculture, cosmetics, and even orthodontics (Cullen et al. 2020 ). Its broad application in applied sciences, one of the fastest-growing fields of scientific research, and technological advancements pique the interest of researchers (Irshad et al. 2021 ; Sher et al. 2021a ; Chakravarty et al. 2022 ). Nanoparticles made of metals such as silver, gold, platinum, nickel, manganese, titanium, and zinc exhibit non-toxicity, antibacterial activity, and catalytic activity. The unique surface plasmon resonance (SPR) properties of gold nanoparticles (AuNPs) make them highly suitable for use in biological fields. They are also easy to produce, can be adjusted in size, and are multifunctional. Furthermore, their properties are well-established. There are many different approaches to synthesis, including chemical and physical techniques; however, it is unfortunate that they all negatively affect the surrounding environment(Lee et al. 2020 ; Hashmi et al. 2021 ). As a result, there is a pressing need to produce metal oxide NPs that are harmless to the environment through simple techniques and plant extracts and diverse biological species(Rasheed and Meera 2016 ). In vitro, the green synthesis of NPs is becoming increasingly popular due to its low environmental impact, low production costs, and high efficiency(Soni et al. 2018 ). This method is easy, rapid, and only requires one step and is suitable for production on a commercial scale. This technique involves converting metal salts into particles and stabilizing them with the use of a biological templates. Metal and metal oxide NPs offer good chemical, physical, and biological capabilities against bacteria, fungus, and viruses due to their high surface area, composition, and shape (Qamar et al. 2021 ; Singh et al. 2021 ). There are numerous methods for producing metal nanoparticles, including chemical and biological processes involving microorganisms and plants. (Singh 2022 ). Due to their extensive surface area, gold nanoparticles (Au NPs) have unique electrical, magnetic, and catalytic properties and vast biological applications (Chen et al. 2022 ; Carrapiço et al. 2023 ; Mitri et al. 2023 ). Au NPs are adaptable and resourceful and can be modified into various shapes depending on the need to switch from one task to another (Manjubaashini and Thangadurai 2023 ). It becomes a strong competitor for silver. These NPs can effectively attack all types of bacteria and viruses compared to silver and other metals and are less hazardous and toxic to the environment when compared to other chemically synthesized nanoparticles (Timoszyk and Grochowalska 2022 ). Because of their resistance to surface oxidation, stability, flexible surface characteristics, and low cytotoxicity, Au NPs are excellent for nanotechnologies such as drug delivery (Patel et al. 2022 ). Grewia asiatica known as phalsa in lower Punjab (Pakistan) is rich in essential nutrients. In the current study Au NPs were produced using Grewia asiatica (locally called phalsa) leaf extract. There have been no studies regarding gold nanoparticle synthesized from this locally occurring phalsa. It is used for instant cooling the body in summer, maintaining electrolyte balance and soothing joint aches, and effectively managing seasonal and chronic conditions(Swain et al. 2023 ). It has a high level of vitamin C, minerals, proteins, phenolics, flavonoids, tannins and anthocyanins. It seeds fruit pulp highly rich in phytochemicals is used to treat different diseases and effective in improving respiratory and cardiac functioning. Their fruit has anticancer, antioxidant, and anti-hyperglycemic characteristics, while the stem bark has analgesic and anti-inflammatory effects(Ranjan et al. 2022 ). Their leaves have antibacterial, anticancer, and antiemetic properties. Hence, the current work is highly motivated to synthesize gold nanoparticles of the smallest possible size using leaf extract of Grewia asiatica (GALE) and to explore the photocatalytic dye degradation, antioxidant and biofilm inhibition properties of synthesized nanoparticles (GAAuNPs). 2. Materials and Methodologies 2.1 Chemicals, Reagents and Plant Source Chemicals utilized in the study were of highest analytical quality and purity. Leaves of the Grewia asiatica (plant) were collected in May from agricultural department of Bahawalpur. The metal used in this experiment were chloroauric acid (HAuCl 4 ·3H 2 O) from Merck. 2,2-diphenyl-1-picrylhydrazyl (DPPH), Sodium Borohydride (NaBH 4 ), Ciprofloxacin and Methylene blue (MB) was purchased from local vendors. De-ionized water used for synthesis or solution preparation. 2.2 Preparation of plant extract GALE was rinsed firstly with simple water and then de-ionized water to remove impurities. The extract of the leaves was prepared by taking 10g of leaf in 100ml of de-ionized water. To get the leaf extract the solution was heated at 90°C in the water bath for 20 minutes with stirring frequently and then allowed to cool at ambient temperature. The extract was taken and filtered. For future usage, the filtered solution was kept in a clean, dry beaker. 2.3 Synthesis of Optimized Gold nanoparticles The 5ml solution of chloroauric acid (1mM) was boiled and 5ml of plant extract was added into it. The mixture was stirred for 2 minutes. The color of the solution appeared as ruby pink. In next 15 minutes, the color changed to violet. The change in color was the indication of preparation of gold nanoparticles. The resulting colloidal solution of AuNPs was then stored for further application and characterization. The steps for synthesizing gold NPs are depicted in Fig. 1 . 2.4 Characterization of synthesized AuNPs The primary methods used for characterization of synthesized nanoparticles include UV-vis spectroscopy, FTIR spectroscopy, SEM, EDX, and X-ray diffraction (XRD). A UV-Vis spectrophotometric investigation was carried out to evaluate the absorbance value of the nanoparticles in the region of 300–600 nm. By measuring SPR, this approach can validate the production of nanomaterials (Mahendran et al. 2022 ). 2.5 Degradation of Organic Dye Plant synthesized GAAuNPs were used as a catalysts at room temperature for catalytic reduction of methylene blue (MB). To begin the catalytic degradation experiment, 2.5ml of MB dye solution (0.04mM) was added to a quartz cuvette with a one-centimeter path length. The dye solution was mixed with 0.5 mL of the newly made NaBH 4 (0.06 M) solution before adding 0.5 mL of colloidal AuNPs. The solution was then gently stirred, and UV absorption was taken at regular intervals. Another sample was created using the same process but without nanoparticles. This was used as a comparison to prove that nanoparticles' reduction was more significant. The following equation was used to measure variations in MB absorption after different intervals to track the dye's reduction reaction: Degradation (%) = 100 (C 0 -C)/C 0 While C = the concentration of dye upon irradiation, C 0 = initial concentration of dye. 2.6 Antibacterial activity We employed Gram-positive ( S. aureus ) and Gram-negative ( E. coli ) bacteria from the University of Agriculture's Institute of Microbiology in Faisalabad, Pakistan, to assess each sample's bactericidal efficacy. Bacteria were cultivated overnight in nutrient agar (Oxoid, UK) at 37°C. Disc diffusion method measured antibacterial activity. Briefly, 100 µL of the investigated microorganism suspension with 10 7 CFU/mL of bacterium cells on a nutrient agar medium was used. Agar plates already inoculated with the tested microorganisms were placed on top of the filter discs (6 mm in diameter), each of which had been individually impregnated with a compound solution. Ciprofloxacin (30 µg/dish) was utilized as a positive reference for bacteria to examine the sensitivity of strains/isolates in the analyzed microbial species. Negative controls were sample-free discs. The plates were placed for 2 hours at 4°C and then incubated at 37°C for 18 hours to accelerate the bacterial growth. Finally, the antibacterial proficiency was assessed by comparing the size of the growth inhibition zones (using a zone reader) (Shahid et al. 2021 ; Shakil et al. 2022 ). 2.7 Biofilm inhibition The biofilm inhibition efficiency of plant extracts and prepared NPs was also assessed and detailed as reported by Perveen et al (Perveen et al. 2021 ). A sterilized 96-well plain plastic tissue cultivation plate was filled with 100 µL of dietary broth, 20 µL of injectable bacterial solution (Oxoid, UK), and 100 µL of the test sample to create the biofilm. Nutrient broth effectively became the sole item in the control wells. After the plates were introduced, they underwent an aerobic incubation at 37 0 C for 24 hours. Each well contents had been thoroughly rinsed three times. The plates were rapidly agitated to remove any germs that were not adhering. The final adhering microbe was consistently detected using 220 microliters of 99% methanol, and the plates were flattened and left to dry for 15 minutes. After 5 minutes, the plates were dyed by pouring 220 mL of 50% crystal violet into each well. To remove the further discoloration, the plates were cleaned. The plates were air-dried; 220 µL of 33% (v/v) glacial acetic acid were added to each well to once again dissolve the dye that had been attached to the adherent cells. At 630 nm, the optical concentration of each well is carefully measured with a micro plate reader (Qasim et al. 2020 ). The findings of the three examinations of each bacterial strain were averaged. The following expression is used to determine the % of inhibition of growth of bacteria. INH % =100 – (OD 630 sample *100)/ OD 630 control 2.8 Antioxidant activity by DPPH assay As described by Siddiqui et al., the radical scavenging capacity of plant extracts and prepared NPs was evaluated using DPPH. (Siddiqui et al. 2020 ). For the antioxidant experiment, 3ml of plant extract was mixed with 1ml of freshly prepared 0.004% DPPH in methanol. The mixture was subsequently placed in a dark area. Then, the absorbance change of the solution was observed at 517 nm. Strong radical scavenging interest exists for reaction aggregates with low absorbance. Additional research was done on the antioxidant activities of butylated hydroxytoluene (BHT) and ascorbic acid. An alternative solution without plant extract was obtained as a control. There have been duplicate runs of every experiment. The IC 50 value was calculated from the graph of inhibition versus sample concentration. 3. Results and Discussion 3.1 UV-visible spectroscopy study of the synthesis of gold nanoparticles The gold nanoparticles were synthesized using the biogenic method in the present study. The formation of synthesized gold nanoparticles was verified by scanning the colloidal solution at various time intervals using a UV-VIS spectrophotometer at wavelengths ranging from 350 to 700 nm. The reduction of metal ions after an interaction with G. asiatica leaf extract (GALE) and the emergence of a ruby red color visibly demonstrate the creation of gold nanoparticles (GAAuNPs). GAAuNPs exhibited two peaks with λ max at 400 nm and 550 nm, as shown in Fig. 2 a. Furthermore, the scanning of the colloidal solution was repeated four times at regular intervals, but no change in absorption spectra appeared, confirming the successful formation of GAAuNPs. Next, the optimized conditions for the synthesis of the GAAuNPs were analyzed. Gold nanoparticles were synthesized using metal salt and extract in 1:1, 1:2, 1:3, and 1:4 ratios. The absorption showed that the optimum concentration for synthesizing GAAuNPs was 1:1 (Fig. 2 b). Similarly, AuNPs were synthesized using different concentrations of precursor metal salt ranging from 1–4 mM. At 1mM concentration of precursor metal salt, the synthesized nanoparticles yielded an intense SPR band, indicating 1mM as the optimum concentration for synthesizing GAAuNPs (Fig. 2 c). Bernawi et al. reported this type of UV-visible absorption spectra for the biosynthesized gold nanoparticle (Barnawi et al. 2022 ). 3.3 FTIR Spectroscopy To analyze the FT-IR spectra of biosynthesized AuNPs, the KBr technique was used. The process involved mixing the AuNPs with KBr under high pressure to create a slice for scanning between 400 and 4000 cm − 1 . The FTIR spectra of leaf extract and produced gold nanoparticles exhibited a broad band at 3374cm − 1 , corresponding to OH group(Sher et al. 2021b ). The weak peaks at 2363cm − 1 , 2068cm − 1 and the strong band at 1650,1380 and 1112 cm − 1 in AuNPs indicate the characteristics of alkynes (C) and carbonyl groups like C = O, COOH groups, respectively (Muniyappan and Nagarajan 2014 ; Thangamani and Bhuvaneshwari 2019 ). These peaks result from the extract's soluble organic components, which may be involved in producing gold nanoparticles (Fig. 3 ). 3.4 X-ray Diffraction Analysis A technique for analyzing the size, dimensions, and composition of nanoparticles is called X-ray diffraction spectroscopy. By using XRD analysis, Au NPs' purity and crystallinity were validated. Figure 4 displays the XRD pattern of GAAuNPs. XRD of GAAuNPs yielded four diffraction peaks at angles of 38.29°, 44.63°, 64.95°, and 77.91° corresponding to diffraction planes of (111), (200), (220), and (311) (Devi et al. 2015 ; Han et al. 2017 ). XRD on Au NPs reveals that the particles have a face-centered cubic structure, which matches the JCPDS card number 04-0784, and the pattern is consistent with previous publications(Kulkarni et al. 2019 ; Ramesh et al. 2019 ). The presence of four peaks specific to Au NPs', as shown in Fig. 5 , indicates the purity of the particles. The Debye-Scherrer equation was used to calculate crystallinity, representing D = kλ/βCosθ where D is the calculated average crystalline size. The average crystallinity of biosynthesized particles was found to be8 nm. 3.5 SEM-EDX analysis The shape and content of GAAuNPs were investigated using scanning electron microscopy combined with energy-dispersive X-rays (SEM-EDX). The GAAuNPs were aggregated and represented in SEM (Fig. 5 a). An EDX analysis was conducted to ascertain the GAAuNPs' elemental makeup. The metallic gold was observed at 2.2 keV, the highest percentage in EDX. In comparison, carbon and oxygen were situated at 0.25 and 0.45 Kev, confirming the organic moiety bound to the gold metal nanoparticle (Fig. 5 b). In EDX spectra, strong Au and O peaks show that gold nanoparticles have formed and are very pure (Fig. 5 b). 3.6 Degradation of Methylene Blue MB, one of the most widely used organic dyes, is harmful in wastewater. in the presence of NaBH4. In the current work, the catalytic performance of biosynthesized gold nanoparticles (GAAuNPs) was checked against MB in the presence of NaBH 4 (reducing agent). A control sample containing NaBH 4 only (without GAAuNPs) was also run to check the catalytic effect of green synthesized AuNPs. A UV-visible spectrophotometer was used to measure changes in MB absorption to track the dye degradation process by gold nanoparticles. Figure 6 b show that NaBH 4 and green-fabricated AuNPs effectively degraded MB by reducing its absorption peak at 664 nm. The presence of Au nanoparticles triggered the breakdown of methylene blue, resulting in complete degradation within 8 minutes. This indicates that the AuNPs were effective in catalyzing the reduction of MB. The control sample (NaBH 4 solution without AuNPs) experienced a slow MB degradation of 15 minutes. Thus, AuNPs acted as excellent catalysts and improved MB degradation (Fig. 6 a). 3.7 Analysis of Biofilm inhibition The GALE and GAAuNPs were subjected to biofilm inhibition assays against S. aureus and E. coli. The GALE showed 42.76% inhibition against E. coli and 7.22% inhibition against S. aureus . In contrast to GALE, the GAAuNPs were more vigorous against both strains and inhibited 55.04% E. coli and 46.12% S. aureus. The biofilm inhibition of GAAuNPs was better than plant extract for both strains as shown in Fig. 7 a. The biosynthesized nanoparticles had an IC 50 of 0.090 against E. coli and 0.10 against S. aureus . These values were similar to the standard (ciprofloxacin) IC 50 values of 0.068 against E. coli and 0.078 against S. aureus (Fig. 7 b). Consequently, it is anticipated that the GAAuNPs that were synthesized will be able to inhibit biofilm formation in both E. coli and S. aureus . Similar results have been reported by previous researchers (Manandhar et al. 2019 ; Hasan et al. 2020 ). 3.8 Antioxidant activity by DPPH assay The antioxidant activities of GALE and synthesized GAAuNPs were tested using the DPPH assay. The results are expressed in IC 50 g/mL, with ascorbic acid serving as the standard. The synthesized GAAuNPs showed more significant antioxidant activity than the standard control (ascorbic acid) (Fig. 8 a). The maximum activity of GAAuNPs may be attributed to the presence of diverse functional groups, as demonstrated in FTIR (Fig. 3 ) (El-Sheekh and El-Kassas 2016 ). The findings suggest that GAAuNPs may be used in place of antioxidants to treat diseases brought on by free radicals. In order to engage with free radicals and convert them into more stable molecules that can halt a chain reaction, GAAuNPs operate as electron donors (Siddiqi et al. 2018 ). AuNPs made from plant extracts exhibit powerful antioxidant capabilities, according to a number of prior investigations (Dorosti and Jamshidi 2016 ; Vimalraj et al. 2018 ). The IC 50 values of leaf extract and AuNPs were 1.24mg/ml and 0.6mg/ml, respectively. Therefore, it is concluded that the biosynthesized GAAuNPs free radical scavenging activity was enhanced compared to the leaf extract. 3.9 Antibacterial Activities Several studies have discovered that AuNPs are more valuable and effective in biological applications than conventional metal NPs (Akintelu et al. 2020 ; Hammami and Alabdallah 2021 ). Because they have a large surface area to interact with the bacterial cell wall, three-dimensionally structured nanoparticles can quickly lead to structural cell deformation (Du et al. 2014 ). The GAAuNPs were subjected to antibacterial activity utilizing bacterial strains; S. aureus and E. coli and ciprofloxacin as a reference standard. The synthesized nanoparticles and leaf extract inhibited two bacterial strains, S. Aureus and E. coli (Fig. 9b). The GALE showed a 12mm and 9mm inhibition zone against E. coli and Staphylococcus , while the GAAuNPs showed a 12mm and 13mm zone of inhibition against both E. coli and Staphylococcus . In this experiment, the GALE had enhanced activity compared to derived nanoparticles. The zone of inhibition for both bacteria was represented graphically in Fig. 9b. The current research has demonstrated that these nanoparticles can combat human diseases because they are effectively absorbed by the peptidoglycan-based cell walls of bacteria. Their small size promotes optimum absorption, ultimately killing the germs (Qamar et al. 2017 ; Khalil et al. 2020 ). According to research, large amounts of hydrogen peroxide and other reactive oxygen species can disrupt protein function by damaging bacterial DNA and cell membranes (Qamar et al. 2022a ; Javed et al. 2023 ). According to the results, nanoparticles are a helpful resource for combating human infections. Conclusions The current work uses GALE to make GAAuNPs straightforwardly and safely. The characterization investigations disclosed the structural aspects and the function of the stabilizing agents during the synthesis of AuNPs. The successful synthesis of AuNPs was verified by a colour shift from light yellowish to ruby red and by scanning via UV-VIS spectroscopy during production. FT-IR investigations have shown the characteristics of alkynes (C), carbonyl groups, and other chemicals in the G. asiatica leaf extract that are responsible for converting Au + 3 ions into GAAuNPs. The SEM-EDX analysis showed nanoparticles that were pure and formed in clusters. The synthetic GAAuNPs used as catalysts showed outstanding methylene blue degradation efficiency. The dye was destroyed in just six minutes. The efficiency of GAAuNPs in degrading the dye was even higher than in previously published studies. The GAAuNPs produced displayed significant biofilm inhibition and antioxidant and antibacterial activity. According to the findings of the study, GAAuNPs can be employed as catalysts and biologically active substances for environmental remediation applications. Declarations Ethics. No ethics approval is necessary since neither humans nor animals were used in the study. Author contributions. S.M.: writing—original draft, formal analysis, methodology; N.A.: investigation, project administration, supervision and validation; M.A.Q: writing—review and editing; A.F, S. N. P: conceptualization, formal analysis, and methodology; A. Y: validation and writing—review and editing; A. N. M. A. A.. Conflict of interest declaration. The authors declare no competing interest. Funding. This work received no funding. References Ahmad N, Javed M, Qamar M A, Kiran U, Shahid S, Akbar M B, Sher M, Amjad A (2022) Synthesis, characterization and potential applications of Ag@ ZnO nanocomposites with S@ gC 3N4. Advances in Materials Research 11(3): 225-235 Akintelu S A, Olugbeko S C, Folorunso A S (2020) A review on synthesis, optimization, characterization and antibacterial application of gold nanoparticles synthesized from plants. 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Catalysts 12(11): 1388 Qamar M A, Shahid S, Javed M, Sher M, Iqbal S, Bahadur A, Li D (2021) Fabricated novel g-C3N4/Mn doped ZnO nanocomposite as highly active photocatalyst for the disinfection of pathogens and degradation of the organic pollutants from wastewater under sunlight radiations. Colloids and Surfaces A: Physicochemical and Engineering Aspects 611: 125863 Qasim N, Shahid M, Yousaf F, Riaz M, Anjum F, Faryad M A, Shabbir R (2020) Therapeutic potential of selected varieties of phoenix dactylifera L. Against microbial biofilm and free radical damage to DNA. Dose-Response 18(4): 1559325820962609 Ramesh A, Tamizhdurai P, Gopinath S, Sureshkumar K, Murugan E, Shanthi K (2019) Facile synthesis of core-shell nanocomposites Au catalysts towards abatement of environmental pollutant Rhodamine B. Heliyon 5(1): e01005 Ranjan P, Brahmi P, Tyagi V, Ranjan J, Srivastava V, Yadav S, Singh S, Singh S, Binda P, Singh S (2022) Global interdependence for fruit genetic resources: status and challenges in India. Food Security 14(3): 591-619 Rasheed R, Meera V (2016) Synthesis of iron oxide nanoparticles coated sand by biological method and chemical method. Procedia Technology 24: 210-216 Shahid M, Naureen I, Riaz M, Anjum F, Fatima H, Rafiq M A (2021) Biofilm inhibition and antibacterial potential of different varieties of garlic (Allium sativum) against sinusitis isolates. Dose-Response 19(4): 15593258211050491 Shakil M, Inayat U, Khalid N, Tanveer M, Gillani S, Tariq N, Shah A, Mahmood A, Dahshan A (2022) Enhanced structural, optical, and photocatalytic activities of Cd–Co doped Zn ferrites for degrading methyl orange dye under irradiation by visible light. Journal of Physics and Chemistry of Solids 161: 110419 Sher M, Javed M, Shahid S, Hakami O, Qamar M A, Iqbal S, Al-Anazy M M, Baghdadi H B (2021a) Designing of highly active g-C3N4/Sn doped ZnO heterostructure as a photocatalyst for the disinfection and degradation of the organic pollutants under visible light irradiation. Journal of Photochemistry and Photobiology A: Chemistry 418: 113393 Sher M, Khan S A, Shahid S, Javed M, Qamar M A, Chinnathambi A, Almoallim H S (2021b) Synthesis of novel ternary hybrid g-C3N4@ Ag-ZnO nanocomposite with Z-scheme enhanced solar light‐driven methylene blue degradation and antibacterial activities. Journal of Environmental Chemical Engineering 9(4): 105366 Siddiqi K S, Husen A, Rao R A (2018) A review on biosynthesis of silver nanoparticles and their biocidal properties. Journal of nanobiotechnology 16(1): 1-28 Siddiqui A J, Danciu C, Ashraf S A, Moin A, Singh R, Alreshidi M, Patel M, Jahan S, Kumar S, Alkhinjar M I (2020) Plants-derived biomolecules as potent antiviral phytomedicines: new insights on ethnobotanical evidences against coronaviruses. Plants 9(9): 1244 Singh A K (2022) A review on plant extract-based route for synthesis of cobalt nanoparticles: Photocatalytic, electrochemical sensing and antibacterial applications. Current Research in Green and Sustainable Chemistry: 100270 Singh K R, Nayak V, Singh J, Singh A K, Singh R P (2021) Potentialities of bioinspired metal and metal oxide nanoparticles in biomedical sciences. RSC advances 11(40): 24722-24746 Singh P, Pandit S, Garnæs J, Tunjic S, Mokkapati V R, Sultan A, Thygesen A, Mackevica A, Mateiu R V, Daugaard A E (2018) Green synthesis of gold and silver nanoparticles from Cannabis sativa (industrial hemp) and their capacity for biofilm inhibition. International journal of nanomedicine: 3571-3591 Soni M, Mehta P, Soni A, Goswami G K (2018) Green nanoparticles: Synthesis and applications. IOSR J. Biotechnol. Biochem 4(3): 78-83 Swain S, Bej S, Mandhata C P, Bishoyi A K, Sahoo C R, Padhy R N (2023) Recent progression on phytochemistry and pharmacological activities of Grewia asiatica L.(Tiliaceae) and traditional uses. South African Journal of Botany 155: 274-287 Swolana D, Kępa M, Idzik D, Dziedzic A, Kabała-Dzik A, Wąsik T J, Wojtyczka R D (2020) The antibacterial effect of silver nanoparticles on Staphylococcus epidermidis strains with different biofilm-forming ability. Nanomaterials 10(5): 1010 Thangamani N, Bhuvaneshwari N (2019) Green synthesis of gold nanoparticles using Simarouba glauca leaf extract and their biological activity of micro-organism. Chemical Physics Letters 732: 136587 Timoszyk A, Grochowalska R (2022) Mechanism and Antibacterial Activity of Gold Nanoparticles (AuNPs) Functionalized with Natural Compounds from Plants. Pharmaceutics 14(12): 2599 Vimalraj S, Ashokkumar T, Saravanan S (2018) Biogenic gold nanoparticles synthesis mediated by Mangifera indica seed aqueous extracts exhibits antibacterial, anticancer and anti-angiogenic properties. Biomedicine & Pharmacotherapy 105: 440-448 Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3798283","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":264755662,"identity":"5875f484-38f9-4920-bbf1-c75ce8d12a59","order_by":0,"name":"Sana Maryam","email":"","orcid":"","institution":"The Government Sadiq College Women University Bahawalpur","correspondingAuthor":false,"prefix":"","firstName":"Sana","middleName":"","lastName":"Maryam","suffix":""},{"id":264755663,"identity":"5f1a8d7c-866b-4945-aa3c-c9a5c3a8a869","order_by":1,"name":"Naseem Akhter","email":"","orcid":"https://orcid.org/0000-0001-8595-015X","institution":"The Government Sadiq College Women University Bahawalpur","correspondingAuthor":false,"prefix":"","firstName":"Naseem","middleName":"","lastName":"Akhter","suffix":""},{"id":264755664,"identity":"432ccad7-c389-4e0f-a4d5-5e2d1ef6743c","order_by":2,"name":"Muhammad Azam Qamar","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzElEQVRIiWNgGAWjYNACA4YEfhCdUECKFskGkBYDEuxJMDgA0UsY8B0/nfi5ouBwnvH51YkfHhgwyPOLHcCvRfJM7mbJMwaHi81uvN0sAXSY4czZCfi1GBzI3SDZYHA4cduNsxtAWhIMbhPScv7t5p8gLZtnnN38gzgtN3K3gW3ZwN+7jThbJG+83WbZYJBeLHGDd5tFgoEEYb/wnc/dfLPhj3Uef//ZzTd/VNjI80sT0MJwAMaQAKuUIKAcRQv/AdyKRsEoGAWjYGQDAA+uS5f7nW28AAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-7645-7133","institution":"University of Management and Technology","correspondingAuthor":true,"prefix":"","firstName":"Muhammad","middleName":"Azam","lastName":"Qamar","suffix":""},{"id":264755665,"identity":"c9e0d92d-25ee-4d23-92eb-941ba0660e0f","order_by":3,"name":"Asma Yaqoob","email":"","orcid":"","institution":"Islamia University: The Islamia University of Bahawalpur Pakistan","correspondingAuthor":false,"prefix":"","firstName":"Asma","middleName":"","lastName":"Yaqoob","suffix":""},{"id":264755666,"identity":"d8e68182-8d45-4d0b-b887-89d79064c0ad","order_by":4,"name":"Muhammad Shahid","email":"","orcid":"","institution":"University of Agriculture Faisalabad","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Shahid","suffix":""},{"id":264755667,"identity":"8f92389f-e4fb-4f60-aa25-ca6e97f1c045","order_by":5,"name":"Ahmad Farhan","email":"","orcid":"","institution":"University of Agriculture Faisalabad","correspondingAuthor":false,"prefix":"","firstName":"Ahmad","middleName":"","lastName":"Farhan","suffix":""},{"id":264755668,"identity":"d4d42424-ca44-47e2-aec0-179cb4027e63","order_by":6,"name":"Shela Parveen Nazir","email":"","orcid":"","institution":"The Government Sadiq College Women University Bahawalpur","correspondingAuthor":false,"prefix":"","firstName":"Shela","middleName":"Parveen","lastName":"Nazir","suffix":""},{"id":264755669,"identity":"0a1a62b3-a006-48de-88e6-e34540bd762b","order_by":7,"name":"Abdel-Nasser M. A. Alaghaz","email":"","orcid":"","institution":"Jazan University","correspondingAuthor":false,"prefix":"","firstName":"Abdel-Nasser","middleName":"M. A.","lastName":"Alaghaz","suffix":""}],"badges":[],"createdAt":"2023-12-24 00:43:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3798283/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3798283/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":49236431,"identity":"22448282-a80a-428f-b5db-1c4ab20fa275","added_by":"auto","created_at":"2024-01-05 17:53:13","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":241894,"visible":true,"origin":"","legend":"\u003cp\u003eSynthesis steps of Au NPs.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/a184023eadcd69465e3326ae.png"},{"id":49237472,"identity":"4bcbf6be-c802-49f1-b670-d38fc25fb789","added_by":"auto","created_at":"2024-01-05 18:01:13","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":163579,"visible":true,"origin":"","legend":"\u003cp\u003eUV-VIS absorption spectra (a) effect of change of metal salt and plant extract ratio (c) effect of change in metal salt concentration on the synthesis of gold nanoparticles using \u003cem\u003eGrewia asiatica\u003c/em\u003eleaf extract\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/0fb271c0c42371cfd2eb0830.png"},{"id":49236429,"identity":"5531f8c9-0f10-4e25-bd85-f5c884896972","added_by":"auto","created_at":"2024-01-05 17:53:13","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":111561,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR analysis of gold nanoparticles synthesized from \u003cem\u003eG.asiatica\u003c/em\u003e extract\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/4c3b6ea3c6df0d8a22033b6a.png"},{"id":49237470,"identity":"a31b3c3c-5feb-4c4f-9360-843b72dba22f","added_by":"auto","created_at":"2024-01-05 18:01:13","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":44231,"visible":true,"origin":"","legend":"\u003cp\u003eSynthesized gold nanoparticles' X-ray diffraction pattern\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/25601f5915245e3e104e2884.png"},{"id":49236433,"identity":"142353fc-3900-4bce-9d46-3982433ee4da","added_by":"auto","created_at":"2024-01-05 17:53:13","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":265320,"visible":true,"origin":"","legend":"\u003cp\u003eSEM (a) EDX analysis of Au nanoparticles (b).\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/44485773b8253106053440d5.png"},{"id":49236435,"identity":"6e63a7b3-a06a-471f-9739-ace1527d511e","added_by":"auto","created_at":"2024-01-05 17:53:13","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":116712,"visible":true,"origin":"","legend":"\u003cp\u003eDegradation of MB in the presence of NaBH\u003csub\u003e4\u003c/sub\u003e (a) and in the presence of NaBH\u003csub\u003e4\u003c/sub\u003e+AuNPs (b).\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/89e8e87141d217c769f8447c.png"},{"id":49237471,"identity":"46191492-1264-4d6c-828a-1aa5541c6352","added_by":"auto","created_at":"2024-01-05 18:01:13","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":44245,"visible":true,"origin":"","legend":"\u003cp\u003eBiofilm inhibition (%) (a) and IC\u003csub\u003e50\u003c/sub\u003e values for Biofilm inhibition by gold nanoparticles (b).\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/a0ae19e2278eb5fff40ba649.png"},{"id":49236436,"identity":"946e2ab4-5323-47b6-8ed6-c4d9ddaad43c","added_by":"auto","created_at":"2024-01-05 17:53:13","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":49395,"visible":true,"origin":"","legend":"\u003cp\u003eDPPH scavenging activity (a) and Antibacterial activity of Au nanoparticles (b)\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/f77860c51fd8a0d6d62c0860.png"},{"id":51157754,"identity":"35bb5ebb-5e76-4468-8c86-8feb9e94e532","added_by":"auto","created_at":"2024-02-15 06:22:10","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1487428,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3798283/v1/6895b740-a8f7-4c21-979f-0477f5c0a9c9.pdf"}],"financialInterests":"","formattedTitle":"Grewia asiatica leaves extract assisted green synthesis of gold nanoparticles and study of their antibacterial, antioxidant and photocatalytic potential","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eDue to the global threat of resistant bacteria, food safety, economic growth, and human and animal health are all at risk (Man et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Puvača et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Additionally, bacteria significantly contribute to persistent and recurring infections because of their propensity to create biofilms, which shield them from the host's immune system, antibiotics, and disinfectants (Singh et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Nassima et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In today's world, it is essential to have an efficient and advanced plan for managing and treating drug-resistant microorganisms (Swolana et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Similarly, an unfavorable ecological shift is accompanied by untreated industrial and domestic effluents being discharged into the ecosystem(Natasha et al. 2020). The waste dye molecules significantly negatively impact humans, wildlife, and vegetation. As a result, organic dye effluents are now recognized as a significant threat to aquatic environments (Iqbal et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Ahmad et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Organic dyes are the most hazardous of all the contaminants released into our environment. The primary contaminants in industrial effluent are dyes because of their color (Javed et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Sher et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2021a\u003c/span\u003e). The biological breakdown of dye molecules is exceedingly difficult because they are tenacious compounds. Due to their toxicity, commercial significance, and environmental effects, dyes have been the subject of extensive research (Qamar et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2022b\u003c/span\u003e). As a result, pathogens and organic contaminants must be removed and detoxified from water bodies more urgently than other pollutants. Different remediation techniques, including chemical, physiochemical, and biological treatments, have so far been suggested. However, each has its pros and cons compared to the others(Sher et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2021a\u003c/span\u003e; Javed et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThese colored pollutants are difficult to remove using traditional methods (such as adsorption, coagulation, flocculation, biodegradation, and so on), and all of these procedures are expensive and require additional planning to remove the byproducts. Alternatively, photocatalytic degradation of hazardous dyes into harmless chemicals is the preferred method for reducing their environmental impact. Developing an effective and long-term solution for disposing of harmful dyes and pathogens in wastewater streams is paramount. Recent advances in nanotechnology have enabled the use of nanoparticles as biological agents for suppressing bacteria and as catalysts for degrading toxic pollutants. Materials at the nanoscale are the focus of nanoscience, which has applications in many different areas, including medicine, engineering, forensics, agriculture, cosmetics, and even orthodontics (Cullen et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Its broad application in applied sciences, one of the fastest-growing fields of scientific research, and technological advancements pique the interest of researchers (Irshad et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Sher et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2021a\u003c/span\u003e; Chakravarty et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eNanoparticles made of metals such as silver, gold, platinum, nickel, manganese, titanium, and zinc exhibit non-toxicity, antibacterial activity, and catalytic activity. The unique surface plasmon resonance (SPR) properties of gold nanoparticles (AuNPs) make them highly suitable for use in biological fields. They are also easy to produce, can be adjusted in size, and are multifunctional. Furthermore, their properties are well-established. There are many different approaches to synthesis, including chemical and physical techniques; however, it is unfortunate that they all negatively affect the surrounding environment(Lee et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Hashmi et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). As a result, there is a pressing need to produce metal oxide NPs that are harmless to the environment through simple techniques and plant extracts and diverse biological species(Rasheed and Meera \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In vitro, the green synthesis of NPs is becoming increasingly popular due to its low environmental impact, low production costs, and high efficiency(Soni et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This method is easy, rapid, and only requires one step and is suitable for production on a commercial scale. This technique involves converting metal salts into particles and stabilizing them with the use of a biological templates. Metal and metal oxide NPs offer good chemical, physical, and biological capabilities against bacteria, fungus, and viruses due to their high surface area, composition, and shape (Qamar et al. \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Singh et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThere are numerous methods for producing metal nanoparticles, including chemical and biological processes involving microorganisms and plants. (Singh \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Due to their extensive surface area, gold nanoparticles (Au NPs) have unique electrical, magnetic, and catalytic properties and vast biological applications (Chen et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Carrapi\u0026ccedil;o et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Mitri et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Au NPs are adaptable and resourceful and can be modified into various shapes depending on the need to switch from one task to another (Manjubaashini and Thangadurai \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). It becomes a strong competitor for silver. These NPs can effectively attack all types of bacteria and viruses compared to silver and other metals and are less hazardous and toxic to the environment when compared to other chemically synthesized nanoparticles (Timoszyk and Grochowalska \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Because of their resistance to surface oxidation, stability, flexible surface characteristics, and low cytotoxicity, Au NPs are excellent for nanotechnologies such as drug delivery (Patel et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). \u003cem\u003eGrewia asiatica\u003c/em\u003e known as phalsa in lower Punjab (Pakistan) is rich in essential nutrients. In the current study Au NPs were produced using \u003cem\u003eGrewia asiatica\u003c/em\u003e (locally called phalsa) leaf extract. There have been no studies regarding gold nanoparticle synthesized from this locally occurring phalsa. It is used for instant cooling the body in summer, maintaining electrolyte balance and soothing joint aches, and effectively managing seasonal and chronic conditions(Swain et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). It has a high level of vitamin C, minerals, proteins, phenolics, flavonoids, tannins and anthocyanins. It seeds fruit pulp highly rich in phytochemicals is used to treat different diseases and effective in improving respiratory and cardiac functioning. Their fruit has anticancer, antioxidant, and anti-hyperglycemic characteristics, while the stem bark has analgesic and anti-inflammatory effects(Ranjan et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Their leaves have antibacterial, anticancer, and antiemetic properties. Hence, the current work is highly motivated to synthesize gold nanoparticles of the smallest possible size using leaf extract of \u003cem\u003eGrewia asiatica\u003c/em\u003e (GALE) and to explore the photocatalytic dye degradation, antioxidant and biofilm inhibition properties of synthesized nanoparticles (GAAuNPs).\u003c/p\u003e"},{"header":"2. Materials and Methodologies","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Chemicals, Reagents and Plant Source\u003c/h2\u003e \u003cp\u003eChemicals utilized in the study were of highest analytical quality and purity. Leaves of the \u003cem\u003eGrewia asiatica\u003c/em\u003e (plant) were collected in May from agricultural department of Bahawalpur. The metal used in this experiment were chloroauric acid (HAuCl\u003csub\u003e4\u003c/sub\u003e\u0026middot;3H\u003csub\u003e2\u003c/sub\u003eO) from Merck. \u003cem\u003e2,2-diphenyl-1-picrylhydrazyl\u003c/em\u003e (DPPH), Sodium Borohydride (NaBH\u003csub\u003e4\u003c/sub\u003e), Ciprofloxacin and Methylene blue (MB) was purchased from local vendors. De-ionized water used for synthesis or solution preparation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Preparation of plant extract\u003c/h2\u003e \u003cp\u003eGALE was rinsed firstly with simple water and then de-ionized water to remove impurities. The extract of the leaves was prepared by taking 10g of leaf in 100ml of de-ionized water. To get the leaf extract the solution was heated at 90\u0026deg;C in the water bath for 20 minutes with stirring frequently and then allowed to cool at ambient temperature. The extract was taken and filtered. For future usage, the filtered solution was kept in a clean, dry beaker.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Synthesis of Optimized Gold nanoparticles\u003c/h2\u003e \u003cp\u003eThe 5ml solution of chloroauric acid (1mM) was boiled and 5ml of plant extract was added into it. The mixture was stirred for 2 minutes. The color of the solution appeared as ruby pink. In next 15 minutes, the color changed to violet. The change in color was the indication of preparation of gold nanoparticles. The resulting colloidal solution of AuNPs was then stored for further application and characterization. The steps for synthesizing gold NPs are depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Characterization of synthesized AuNPs\u003c/h2\u003e \u003cp\u003eThe primary methods used for characterization of synthesized nanoparticles include UV-vis spectroscopy, FTIR spectroscopy, SEM, EDX, and X-ray diffraction (XRD). A UV-Vis spectrophotometric investigation was carried out to evaluate the absorbance value of the nanoparticles in the region of 300\u0026ndash;600 nm. By measuring SPR, this approach can validate the production of nanomaterials (Mahendran et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Degradation of Organic Dye\u003c/h2\u003e \u003cp\u003ePlant synthesized GAAuNPs were used as a catalysts at room temperature for catalytic reduction of methylene blue (MB). To begin the catalytic degradation experiment, 2.5ml of MB dye solution (0.04mM) was added to a quartz cuvette with a one-centimeter path length. The dye solution was mixed with 0.5 mL of the newly made NaBH\u003csub\u003e4\u003c/sub\u003e (0.06 M) solution before adding 0.5 mL of colloidal AuNPs. The solution was then gently stirred, and UV absorption was taken at regular intervals. Another sample was created using the same process but without nanoparticles. This was used as a comparison to prove that nanoparticles' reduction was more significant. The following equation was used to measure variations in MB absorption after different intervals to track the dye's reduction reaction:\u003c/p\u003e \u003cp\u003eDegradation (%)\u0026thinsp;=\u0026thinsp;100 (C\u003csub\u003e0\u003c/sub\u003e -C)/C\u003csub\u003e0\u003c/sub\u003e\u003c/p\u003e \u003cp\u003eWhile C\u0026thinsp;=\u0026thinsp;the concentration of dye upon irradiation, C\u003csub\u003e0\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;initial concentration of dye.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e\u003cb\u003e2.6 Antibacterial activity\u003c/b\u003e\u003c/h2\u003e \u003cp\u003eWe employed Gram-positive (\u003cem\u003eS. aureus\u003c/em\u003e) and Gram-negative (\u003cem\u003eE. coli\u003c/em\u003e) bacteria from the University of Agriculture's Institute of Microbiology in Faisalabad, Pakistan, to assess each sample's bactericidal efficacy. Bacteria were cultivated overnight in nutrient agar (Oxoid, UK) at 37\u0026deg;C. Disc diffusion method measured antibacterial activity. Briefly, 100 \u0026micro;L of the investigated microorganism suspension with 10\u003csup\u003e7\u003c/sup\u003e CFU/mL of bacterium cells on a nutrient agar medium was used. Agar plates already inoculated with the tested microorganisms were placed on top of the filter discs (6 mm in diameter), each of which had been individually impregnated with a compound solution. Ciprofloxacin (30 \u0026micro;g/dish) was utilized as a positive reference for bacteria to examine the sensitivity of strains/isolates in the analyzed microbial species. Negative controls were sample-free discs. The plates were placed for 2 hours at 4\u0026deg;C and then incubated at 37\u0026deg;C for 18 hours to accelerate the bacterial growth. Finally, the antibacterial proficiency was assessed by comparing the size of the growth inhibition zones (using a zone reader) (Shahid et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Shakil et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Biofilm inhibition\u003c/h2\u003e \u003cp\u003eThe biofilm inhibition efficiency of plant extracts and prepared NPs was also assessed and detailed as reported by Perveen et al (Perveen et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). A sterilized 96-well plain plastic tissue cultivation plate was filled with 100 \u0026micro;L of dietary broth, 20 \u0026micro;L of injectable bacterial solution (Oxoid, UK), and 100 \u0026micro;L of the test sample to create the biofilm. Nutrient broth effectively became the sole item in the control wells. After the plates were introduced, they underwent an aerobic incubation at 37\u003csup\u003e0\u003c/sup\u003eC for 24 hours. Each well contents had been thoroughly rinsed three times. The plates were rapidly agitated to remove any germs that were not adhering. The final adhering microbe was consistently detected using 220 microliters of 99% methanol, and the plates were flattened and left to dry for 15 minutes. After 5 minutes, the plates were dyed by pouring 220 mL of 50% crystal violet into each well. To remove the further discoloration, the plates were cleaned. The plates were air-dried; 220 \u0026micro;L of 33% (v/v) glacial acetic acid were added to each well to once again dissolve the dye that had been attached to the adherent cells. At 630 nm, the optical concentration of each well is carefully measured with a micro plate reader (Qasim et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The findings of the three examinations of each bacterial strain were averaged. The following expression is used to determine the % of inhibition of growth of bacteria.\u003c/p\u003e \u003cp\u003eINH % =100 \u0026ndash; (OD \u003csub\u003e630 sample\u003c/sub\u003e *100)/ OD \u003csub\u003e630 control\u003c/sub\u003e\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.8 Antioxidant activity by DPPH assay\u003c/h2\u003e \u003cp\u003eAs described by Siddiqui et al., the radical scavenging capacity of plant extracts and prepared NPs was evaluated using DPPH. (Siddiqui et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). For the antioxidant experiment, 3ml of plant extract was mixed with 1ml of freshly prepared 0.004% DPPH in methanol. The mixture was subsequently placed in a dark area. Then, the absorbance change of the solution was observed at 517 nm. Strong radical scavenging interest exists for reaction aggregates with low absorbance. Additional research was done on the antioxidant activities of butylated hydroxytoluene (BHT) and ascorbic acid. An alternative solution without plant extract was obtained as a control. There have been duplicate runs of every experiment. The IC\u003csub\u003e50\u003c/sub\u003e value was calculated from the graph of inhibition versus sample concentration.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.1 UV-visible spectroscopy study of the synthesis of gold nanoparticles\u003c/h2\u003e \u003cp\u003eThe gold nanoparticles were synthesized using the biogenic method in the present study. The formation of synthesized gold nanoparticles was verified by scanning the colloidal solution at various time intervals using a UV-VIS spectrophotometer at wavelengths ranging from 350 to 700 nm. The reduction of metal ions after an interaction with \u003cem\u003eG. asiatica\u003c/em\u003e leaf extract (GALE) and the emergence of a ruby red color visibly demonstrate the creation of gold nanoparticles (GAAuNPs). GAAuNPs exhibited two peaks with λ\u003csub\u003emax\u003c/sub\u003e at 400 nm and 550 nm, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea. Furthermore, the scanning of the colloidal solution was repeated four times at regular intervals, but no change in absorption spectra appeared, confirming the successful formation of GAAuNPs. Next, the optimized conditions for the synthesis of the GAAuNPs were analyzed. Gold nanoparticles were synthesized using metal salt and extract in 1:1, 1:2, 1:3, and 1:4 ratios. The absorption showed that the optimum concentration for synthesizing GAAuNPs was 1:1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb). Similarly, AuNPs were synthesized using different concentrations of precursor metal salt ranging from 1\u0026ndash;4 mM. At 1mM concentration of precursor metal salt, the synthesized nanoparticles yielded an intense SPR band, indicating 1mM as the optimum concentration for synthesizing GAAuNPs (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec). Bernawi et al. reported this type of UV-visible absorption spectra for the biosynthesized gold nanoparticle (Barnawi et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.3 FTIR Spectroscopy\u003c/h2\u003e \u003cp\u003eTo analyze the FT-IR spectra of biosynthesized AuNPs, the KBr technique was used. The process involved mixing the AuNPs with KBr under high pressure to create a slice for scanning between 400 and 4000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The FTIR spectra of leaf extract and produced gold nanoparticles exhibited a broad band at 3374cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, corresponding to OH group(Sher et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2021b\u003c/span\u003e). The weak peaks at 2363cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 2068cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and the strong band at 1650,1380 and 1112 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e in AuNPs indicate the characteristics of alkynes (C) and carbonyl groups like C\u0026thinsp;=\u0026thinsp;O, COOH groups, respectively (Muniyappan and Nagarajan \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Thangamani and Bhuvaneshwari \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). These peaks result from the extract's soluble organic components, which may be involved in producing gold nanoparticles (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.4 X-ray Diffraction Analysis\u003c/h2\u003e \u003cp\u003eA technique for analyzing the size, dimensions, and composition of nanoparticles is called X-ray diffraction spectroscopy. By using XRD analysis, Au NPs' purity and crystallinity were validated. Figure\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e displays the XRD pattern of GAAuNPs. XRD of GAAuNPs yielded four diffraction peaks at angles of 38.29\u0026deg;, 44.63\u0026deg;, 64.95\u0026deg;, and 77.91\u0026deg; corresponding to diffraction planes of (111), (200), (220), and (311) (Devi et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Han et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). XRD on Au NPs reveals that the particles have a face-centered cubic structure, which matches the JCPDS card number 04-0784, and the pattern is consistent with previous publications(Kulkarni et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Ramesh et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The presence of four peaks specific to Au NPs', as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, indicates the purity of the particles. The Debye-Scherrer equation was used to calculate crystallinity, representing D\u0026thinsp;=\u0026thinsp;kλ/βCosθ where D is the calculated average crystalline size. The average crystallinity of biosynthesized particles was found to be8 nm.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e3.5 SEM-EDX analysis\u003c/h2\u003e \u003cp\u003eThe shape and content of GAAuNPs were investigated using scanning electron microscopy combined with energy-dispersive X-rays (SEM-EDX). The GAAuNPs were aggregated and represented in SEM (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003ea). An EDX analysis was conducted to ascertain the GAAuNPs' elemental makeup. The metallic gold was observed at 2.2 keV, the highest percentage in EDX. In comparison, carbon and oxygen were situated at 0.25 and 0.45 Kev, confirming the organic moiety bound to the gold metal nanoparticle (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eb). In EDX spectra, strong Au and O peaks show that gold nanoparticles have formed and are very pure (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e3.6 Degradation of Methylene Blue\u003c/h2\u003e \u003cp\u003eMB, one of the most widely used organic dyes, is harmful in wastewater. in the presence of NaBH4. In the current work, the catalytic performance of biosynthesized gold nanoparticles (GAAuNPs) was checked against MB in the presence of NaBH\u003csub\u003e4\u003c/sub\u003e (reducing agent). A control sample containing NaBH\u003csub\u003e4\u003c/sub\u003e only (without GAAuNPs) was also run to check the catalytic effect of green synthesized AuNPs. A UV-visible spectrophotometer was used to measure changes in MB absorption to track the dye degradation process by gold nanoparticles. Figure\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eb show that NaBH\u003csub\u003e4\u003c/sub\u003e and green-fabricated AuNPs effectively degraded MB by reducing its absorption peak at 664 nm. The presence of Au nanoparticles triggered the breakdown of methylene blue, resulting in complete degradation within 8 minutes. This indicates that the AuNPs were effective in catalyzing the reduction of MB. The control sample (NaBH\u003csub\u003e4\u003c/sub\u003e solution without AuNPs) experienced a slow MB degradation of 15 minutes. Thus, AuNPs acted as excellent catalysts and improved MB degradation (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ea).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e3.7 Analysis of Biofilm inhibition\u003c/h2\u003e \u003cp\u003eThe GALE and GAAuNPs were subjected to biofilm inhibition assays against \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eE. coli.\u003c/em\u003e The GALE showed 42.76% inhibition against \u003cem\u003eE. coli\u003c/em\u003e and 7.22% inhibition against \u003cem\u003eS. aureus\u003c/em\u003e. In contrast to GALE, the GAAuNPs were more vigorous against both strains and inhibited 55.04% \u003cem\u003eE. coli\u003c/em\u003e and 46.12% \u003cem\u003eS. aureus.\u003c/em\u003e The biofilm inhibition of GAAuNPs was better than plant extract for both strains as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003ea. The biosynthesized nanoparticles had an IC\u003csub\u003e50\u003c/sub\u003e of 0.090 against \u003cem\u003eE. coli\u003c/em\u003e and 0.10 against \u003cem\u003eS. aureus\u003c/em\u003e. These values were similar to the standard (ciprofloxacin) IC\u003csub\u003e50\u003c/sub\u003e values of 0.068 against \u003cem\u003eE. coli\u003c/em\u003e and 0.078 against \u003cem\u003eS. aureus\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eb). Consequently, it is anticipated that the GAAuNPs that were synthesized will be able to inhibit biofilm formation in both \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e. Similar results have been reported by previous researchers (Manandhar et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Hasan et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e3.8 Antioxidant activity by DPPH assay\u003c/h2\u003e \u003cp\u003eThe antioxidant activities of GALE and synthesized GAAuNPs were tested using the DPPH assay. The results are expressed in IC\u003csub\u003e50\u003c/sub\u003e g/mL, with ascorbic acid serving as the standard. The synthesized GAAuNPs showed more significant antioxidant activity than the standard control (ascorbic acid) (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003ea). The maximum activity of GAAuNPs may be attributed to the presence of diverse functional groups, as demonstrated in FTIR (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) (El-Sheekh and El-Kassas \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The findings suggest that GAAuNPs may be used in place of antioxidants to treat diseases brought on by free radicals. In order to engage with free radicals and convert them into more stable molecules that can halt a chain reaction, GAAuNPs operate as electron donors (Siddiqi et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). AuNPs made from plant extracts exhibit powerful antioxidant capabilities, according to a number of prior investigations (Dorosti and Jamshidi \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Vimalraj et al. \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The IC\u003csub\u003e50\u003c/sub\u003e values of leaf extract and AuNPs were 1.24mg/ml and 0.6mg/ml, respectively. Therefore, it is concluded that the biosynthesized GAAuNPs free radical scavenging activity was enhanced compared to the leaf extract.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003e3.9 Antibacterial Activities\u003c/h2\u003e \u003cp\u003eSeveral studies have discovered that AuNPs are more valuable and effective in biological applications than conventional metal NPs (Akintelu et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Hammami and Alabdallah \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Because they have a large surface area to interact with the bacterial cell wall, three-dimensionally structured nanoparticles can quickly lead to structural cell deformation (Du et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The GAAuNPs were subjected to antibacterial activity utilizing bacterial strains; \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e and ciprofloxacin as a reference standard. The synthesized nanoparticles and leaf extract inhibited two bacterial strains, \u003cem\u003eS. Aureus\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e (Fig.\u0026nbsp;9b). The GALE showed a 12mm and 9mm inhibition zone against \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eStaphylococcus\u003c/em\u003e, while the GAAuNPs showed a 12mm and 13mm zone of inhibition against both \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eStaphylococcus\u003c/em\u003e. In this experiment, the GALE had enhanced activity compared to derived nanoparticles. The zone of inhibition for both bacteria was represented graphically in Fig.\u0026nbsp;9b. The current research has demonstrated that these nanoparticles can combat human diseases because they are effectively absorbed by the peptidoglycan-based cell walls of bacteria. Their small size promotes optimum absorption, ultimately killing the germs (Qamar et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Khalil et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). According to research, large amounts of hydrogen peroxide and other reactive oxygen species can disrupt protein function by damaging bacterial DNA and cell membranes (Qamar et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022a\u003c/span\u003e; Javed et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). According to the results, nanoparticles are a helpful resource for combating human infections.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe current work uses GALE to make GAAuNPs straightforwardly and safely. The characterization investigations disclosed the structural aspects and the function of the stabilizing agents during the synthesis of AuNPs. The successful synthesis of AuNPs was verified by a colour shift from light yellowish to ruby red and by scanning via UV-VIS spectroscopy during production. FT-IR investigations have shown the characteristics of alkynes (C), carbonyl groups, and other chemicals in the \u003cem\u003eG. asiatica\u003c/em\u003e leaf extract that are responsible for converting Au\u003csup\u003e+\u0026thinsp;3\u003c/sup\u003e ions into GAAuNPs. The SEM-EDX analysis showed nanoparticles that were pure and formed in clusters. The synthetic GAAuNPs used as catalysts showed outstanding methylene blue degradation efficiency. The dye was destroyed in just six minutes. The efficiency of GAAuNPs in degrading the dye was even higher than in previously published studies. The GAAuNPs produced displayed significant biofilm inhibition and antioxidant and antibacterial activity. According to the findings of the study, GAAuNPs can be employed as catalysts and biologically active substances for environmental remediation applications.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics.\u0026nbsp;\u003c/strong\u003eNo ethics approval is necessary since neither humans nor animals were used in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions.\u0026nbsp;\u003c/strong\u003eS.M.: writing\u0026mdash;original draft, formal analysis, methodology; N.A.: investigation, project administration, supervision and validation; M.A.Q: writing\u0026mdash;review and editing; A.F, S. N. P: conceptualization, formal analysis, and methodology; A. Y: validation and writing\u0026mdash;review and editing; A. N. M. A. A..\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest declaration.\u0026nbsp;\u003c/strong\u003eThe authors declare no competing interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding.\u0026nbsp;\u003c/strong\u003eThis work received no funding.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAhmad N, Javed M, Qamar M A, Kiran U, Shahid S, Akbar M B, Sher M, Amjad A (2022) Synthesis, characterization and potential applications of Ag@ ZnO nanocomposites with S@ gC 3N4. Advances in Materials Research 11(3): 225-235\u003c/li\u003e\n\u003cli\u003eAkintelu S A, Olugbeko S C, Folorunso A S (2020) A review on synthesis, optimization, characterization and antibacterial application of gold nanoparticles synthesized from plants. 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Biomedicine \u0026amp; Pharmacotherapy 105: 440-448\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Green nanoparticles, dye degradation, biofilm inhibition, antibacterial activity","lastPublishedDoi":"10.21203/rs.3.rs-3798283/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3798283/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe environmental issues generated by industrial advancement and human activities are serious and must not be overlooked. Bio-nanotechnology has emerged as a solution for producing biosynthetic nanomaterials to address environmental issues. The current work describes for the first time an economical and environmentally benign method of producing gold nanoparticles utilizing \u003cem\u003eGrewia asiatica\u003c/em\u003e (GAAuNPs). The optimal AuNPs were produced using a gold ion concentration of 1 mM and a metal-to-extract ratio of 1:1. The resulting nanoparticles were analyzed and characterized using X-ray crystallography, FTIR, SEM, EDX, and UV visible spectroscopy. The catalytic methylene blue (MB) dye degradation properties, antibacterial and biofilm inhibition abilities, and antioxidant properties of the synthesized particles were investigated. The Au NPs demonstrated efficient catalytic degradation against MB and completely degraded it in 8 minutes. The NPs also exhibited potent biofilm inhibition against \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e that was close to the standard and was more antioxidant than the standard. The findings highlight \u003cem\u003eG. asiatica\u003c/em\u003e as a suitable, inexpensive biosource for biofabricating GAAuNPs with extensive multifunctional uses. To conclude, GAAuNPs' ability to degrade organic pollutants and eradicate pathogens provides a cost-effective and environmentally benign remedy to tackle contemporary pollutants.\u003c/p\u003e","manuscriptTitle":"Grewia asiatica leaves extract assisted green synthesis of gold nanoparticles and study of their antibacterial, antioxidant and photocatalytic potential","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-05 17:53:08","doi":"10.21203/rs.3.rs-3798283/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"758ebbef-d8d1-4708-9a7f-e43fe26c81f8","owner":[],"postedDate":"January 5th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-02-15T06:21:57+00:00","versionOfRecord":[],"versionCreatedAt":"2024-01-05 17:53:08","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3798283","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3798283","identity":"rs-3798283","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}