Optimization on production and technological properties of exopolysaccharide from Pediococcus pentosaceus SL4 isolated from sourdough Tomi-tomi (Flacourtia Inermis Roxb)

Optimization on production and technological properties of exopolysaccharide from Pediococcus pentosaceus SL4 isolated from sourdough Tomi-tomi (Flacourtia Inermis Roxb) | 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 Optimization on production and technological properties of exopolysaccharide from Pediococcus pentosaceus SL4 isolated from sourdough Tomi-tomi (Flacourtia Inermis Roxb) Dyah Ayu Puspitasari, Rini Yanti, Chusnul Hidayat, Dian Anggraini Suroto This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7555681/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 25 Mar, 2026 Read the published version in World Journal of Microbiology and Biotechnology → Version 1 posted 10 You are reading this latest preprint version Abstract Sourdough is a fermented food made by a complex and diverse consortium of microorganisms, predominantly wild yeasts and lactic acid bacteria (LAB), positioning it as a valuable source for the isolation of specific LAB strains. LAB are capable of synthesising metabolites known as exopolysaccharides (EPS) during fermentation which enhanced the quality of food product. The characteristics of exopolysaccharides which are produced by LAB vary greatly depending on structure, molecular size, and physicochemical properties. This study aims to determine the optimal conditions for EPS production and characterize the exopolysaccharide from Pediococcus pentosaceus SL4 isolated from Flacourtia inermis sourdough. The investigation utilized a modified de Man Sharpe Rogosa medium and response surface methodology (RSM). The screened factors included monosaccharide types and concentrations, inoculum size, agitation speeds, and fermentation time. Using the Box Behnken design, the optimized conditions for EPS production were sucrose concentration of 5%, inoculum size of 6%, and agitation at 100 rpm, resulting in a maximum yield of 426.2 mg/L after 24 h incubation. The partially purified EPS had a molecular weight of 7.35×10 5 kDa with a degradation temperature of 91.8⁰C. The irregular, porous, and spongy morphology of EPS was observed by scanning electron microscopy. The EPS exhibited physiological properties such as water solubility index, holding capacity, and emulsifying activity. Overall, EPS exhibits technological properties as a stabilizer in the food industry. Box Behnken design characterization exopolysaccharides food stabilizer P. pentosaceus SL4 Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction The demand for biomolecules that offer technological and health benefits has been increasing and must be addressed by exploring alternative biomolecule sources with specific properties. Exopolysaccharides (EPS) are a type of extracellular polysaccharide produced by bacteria that have wide applications in the food, cosmetic and pharmaceutical industries, functioning as a stabilizer, texturizer, emulsifier, and gelling agent (Wang et al. 2020 ; Balyan et al. 2024a ). Some groups of lactic acid bacteria (LAB), such as Lactobacillus , Enterococcus , and Pediococcus , produce EPS through sucrose supplementation in the fermentation medium (Jayamanohar et al. 2018 ; Wang et al. 2019 ; Jiang et al. 2022 ) to protect themselves from environmental stresses such as insufficient nutrients, pH, salinity, temperature, or co-cultivation (Wang et al. 2017 ; Vinothini et al. 2019 ; Oleksy-Sobczak and Klewicka 2020 ; Jiang et al. 2020 ). Depending on the sugar composition, EPS can be either homopolysaccharides or heteropolysaccharides. The properties of EPS are associated with their characteristics, such as molecular weight, thermal properties, and sugar composition. EPS produced by LAB has the potential to play an important role because it has been classified as Generally Recognized as Safe (GRAS). The properties of EPS vary depending on the strain and fermentation conditions. Consequently, screening for LAB shows potential EPS production is necessary. Fermented foodstuffs like sourdough could be an ideal source for EPS-producing LAB. LABs exist spontaneously in sourdough due to the materials utilized or the environment where the sourdough is produced (Ripari et al. 2016 ; Yu et al. 2018 ). Some LABs, including P. pentosaceus , L. plantarum , and L. brevis , have been isolated from sourdough and produce EPS (Ripari et al., 2016 ; Fu et al., 2022 ). The presence of exopolysaccharides produced by LAB has been shown to improve the rheological properties of sourdough (Zhang et al. 2023a ) and enhance health benefits such as antioxidants (Wang et al. 2020 ), maintain blood sugar stability, and reduce cholesterol (Dilna et al. 2015 ). Other studies reported that the utilization of lactic acid bacteria, such as Lactobacillus spp., Pediococcus spp., and Leuconostoc spp ., in the production of EPS has been demonstrated to enhance the stability and quality of food products through its application as a stabilizer and a coating agent (Wang et al, 2019 ; Jiang et al. 2022 ; Balyan et al. 2024). We developed the sourdough using some tropical fruits as a starter (Kemhay et al. 2024 ) and we also isolated Pediococccus pentosaceus SL4 were among the LAB species and had been screened for its potential EPS production (unpublished data). P. pentosaceus cannot produce large amounts of exopolysaccharides without optimal conditions, while under optimal conditions, LAB produces EPS that can be 2 to 5 times higher depending on the bacterial strain (Xing et al. 2018 ; Wang et al. 2019 ; Jiang et al. 2020 ). The optimization of carbon sources and fermentation process conditions has been identified as a strategy to enhance EPS production (Oleksy-Sobczak et al. 2020 ; Zanzan et al. 2023 ). However, it is crucial to note that each strain exhibits distinct optimal conditions for EPS production. The objectives of this study are to optimize EPS production of Pediococccus pentosaceus SL4 by adjusting carbon source, agitation speed, inoculum size, and fermentation time and subsequently derive a quadratic model to biosynthesize EPS and to characterize the EPS using various instrumental analyses and estimate its technological properties as a stabilizer in food processing. 2. Materials and Methods Materials The high-yield EPS-producing strain P. pentosaceus SL4 was isolated from tomi-tomi sourdough and was stored at -80⁰C in the Biotechnology Laboratory Faculty of Agricultural Technology Gadjah Mada University. de Man Rogosa Sharpe (MRS) broth (Himedia, India) supplemented with saccharides was used to cultivate the SL4 for EPS production. Glucose, fructose, galactose, and lactose as supplemented sugar were from Merck (Darmstadt, Germany) and sucrose from Phytotech (Kansas, USA). All chemicals used for analysis were analytic grade from Sigma Aldrich (Missouri, USA), and virgin red palm oil was from the local Indonesian market. Preliminary screening of fermentation variables and optimization Screening design is a methodical approach that facilitates the comprehension of the factors potentially implicated in the system. The bacteria were inoculated in MRS broth for 18 hours at 37°C for pre-culture. The first stage in screening was determining the right carbon source. Five types of carbon sources at 20 g/L were supplemented to MRS broth, culture at concentration 2% (v/v) was inoculated into sterile broth, and fermentation was carried out at 37⁰C with 80 rpm agitation for 48 hours. The best carbon source screening results were used for the next screening stage. In the second stage, the variables utilized were sucrose (25–125 g/L), inoculum size (2–10%), agitation speeds (0-200 rpm), and cultivation time (8–56 h) used to evaluate the maximum EPS production. The starter was inoculated at specific conditions based on one factor at a time (OFAT) with four numerical factors. After identifying the factors affecting EPS production by OFAT design, a Box-Behnken design (BBD) was employed to assess the impact of three additional factors. The 2% culture (v/v) was inoculated to MRS broth (5% glucose) and incubated at 80 rpm at 37⁰C for 48 h, which was used as a non-optimized fermentation condition. Extraction and purification of EPS The extraction of exopolysaccharide (EPS) from P. pentosaceus was performed according to the method of Zanzan et al. ( 2023 ) with modifications. After incubation, the cultures were heated at 100°C for 10 minutes to inactivate EPS degrading enzymes, followed by centrifugation at 5000×g for 30 minutes. The pre-chilled 96% (v/v) ethanol was added to the resultant precipitate twice the initial volume. The mixture was then stored at 4°C for 24 hours, followed by precipitation using centrifugation at 5000×g for 30 minutes at a temperature of 4°C. Subsequently, 8% (w/v) trichloroacetic acid (TCA) was added to the precipitates and maintained at 4°C for 30 minutes to yield a protein precipitate. The protein precipitates were then removed by centrifugation at 5000 × g for 30 minutes. Subsequently, two volumes of pre-chilled 96% (v/v) ethanol were added to the filtrate and stored at 4°C for 24 hours, and the precipitate was subjected to centrifugation at 5000×g for 30 minutes at 4°C. The polysaccharide pellet was dissolved in 5 mL of deionized water and dialyzed using a 12–14 kDa cut-off dialysis membrane. The resulting EPS was then lyophilized, resulting in a powder form of the EPS suitable for further analysis. Characterization of EPS Molecular weight analysis Size exclusion chromatography was used to determine the molecular weight distribution of EPS. Ten milligrams of EPS sample were dissolved in 1 mL of ddH 2 O and filtered through a 0.22 µm filter. Then, the EPS solution (10 mg/mL, 20µL) was loaded into the gel permeation chromatography system, TOSOH HLC-8320GPC Japan, with TSKGel Super AW5000 as column and coupled to the Refractive Index detector. Isocratic elution was carried out using deionized water at a constant rate of 0.3 mL/min, and the detector temperature was set to 40⁰C. Fourier transform infrared (FT-IR) analysis Ten milligrams (10 mg) of the lyophilized EPS sample were ground with KBr powder. The spectrum was recorded using a Bruker vertex 70 DTGS FTIR spectrometer (Bruker, Germany) with a resolution of 4 cm -1 in the 400–4000 cm -1 range. Scanning Electron Microscopy (SEM) and Energy-dispersive X-ray analysis of EPS The morphology of lyophilized EPS was observed under SEM JEOL JSM-IT200 (Tokyo, Japan). In brief, 5 mg of lyophilized EPS was placed on SEM stubs, tapped with double-sided, and coated with a carbon layer of approximately 10 nm thickness. The image has been taken at 10 kV at a resolution of 250×, 2000×, and 15000× magnification. The same samples for SEM were used for EDX analysis but with an acceleration voltage of 20 kV and at 250x magnification. Thermal properties Differential scanning calorimetry (NETZSCH DSC 300 Caliris Select, Germany) was used to investigate the thermal properties of EPS. The EPS sample (3–5 mg) was put in an alumina pan and sealed. The system was heated from 25⁰C to 400⁰C at a linear heating rate of 10⁰C/min in a nitrogen atmosphere (50 mL/min). The empty aluminium crucible was treated as a reference. Water Holding Capacity Water holding capacity (WHC) was determined by dissolving 10 mg of EPS in 1 mL of double distilled water and mixing in the vortex for 5 min. The sample was then centrifuged at 16000 rpm for 25 min. The pellet was collected and weighed (Marimuthu and Rajendran 2023 ). The WHC was determined using Eq. (1) Solubility Index The EPS sample (10 mg) was dissolved in 1 mL double distilled water under constant agitation for 24 h, then centrifuged at 6000 rpm for 20 min. A triple volume of chilled ethanol 96% was added to the supernatant to precipitate EPS. It was then centrifuged at 12000 rpm for 20 min. The pellet was dried at 55⁰C and weighed (Marimuthu and Rajendran 2023 ). The solubility was calculated using Eq. (2) Emulsification properties of EPS The emulsion index was evaluated by suspended EPS at various concentrations (0.25–1.25 mg/mL) in double distilled water as described previously (Marimuthu and Rajendran 2023 ) with slight modification. A 2 mL of EPS solution was added to an equal volume of virgin red palm oil, and the mixture was vortexed vigorously for 5 min. The emulsion activity (%EA) was defined as the percent retention of the emulsion after incubation for one hour at room temperature, whereas emulsion stability (%ES) was determined after 24, 48, and 72 h separately. The %EA and %ES calculated according to Eq. (3) 3. Results Effect of media composition and culture conditions on EPS production A comprehensive experimental approach was employed to ascertain the optimal carbon source for enhancing exopolysaccharide production. This involved utilizing five distinct carbon sources, each at a concentration of 20 g/L, within a customized MRS broth medium (Table 1). The experimental design comprised a one-factor-at-a-time (OFAT) experimental framework meticulously designed to investigate the impact of medium composition and prevailing culture conditions on the production of exopolysaccharides. No statistically significant differences (p>0.05) in EPS yield or cell growth were observed when five carbon sources were prepared as supplements in fermentation media. However, sucrose exhibited the highest EPS production at 85 mg/L, 1.5-fold higher than MRS without saccharide supplementation. After determining sucrose as the optimal carbon source, different sucrose concentrations (25–125 g/L) were examined to ascertain the appropriate sucrose concentration for EPS production. The preliminary screening revealed optimal EPS production was achieved at 150.1 mg/L with 50 g/L sucrose supplementation. [Table 1] A culture was also cultivated on a water bath shaker to ascertain the impact of agitation on EPS production. The agitation speed of the water bath shaker was adjusted within the range of 0 to 150 rpm and the quantity of produced EPS was examined after 48 hours. The application of an agitation speed of 100 rpm was found to enhance EPS production by up to 2-fold compared to fermentation conducted without agitation. A statistically significant difference (p<0.05) was observed between cultures cultivated with and without agitation (Figure 1). Agitation speeds exceeding 100 rpm during fermentation did not lead to a significant enhancement in EPS production. A preliminary study was conducted to evaluate the effect of inoculum size on EPS production using inoculum concentrations of 2%, 4%, 6%, 8%, and 10%. Fermentation was carried out at an agitation speed of 100 rpm, and EPS was harvested after 48 hours. The results indicated that the highest EPS yield was obtained with an inoculum size of 6%, reaching 204.9 mg/L—approximately 1.2 times higher than that obtained with lower concentrations. Increasing the inoculum size beyond 6% did not result in a significant enhancement of EPS production. [Figure 1] Fermentation time is critical in producing exopolysaccharides (EPS) due to different microorganisms varying growth phase rates. In this screening stage, the determination of the optimal fermentation time was carried out by culturing with a 6% starter concentration, 50 g/L sucrose supplementation, and an agitation speed of 100 rpm for 8, 16, 24, 32, 40, 48, and 56 hours. The results showed that the highest EPS production was achieved at 24 hours of fermentation. Extending the fermentation time did not result in a further increase in the number of EPS produced. Optimization and validation of EPS yield Following the implementation of a screening process employing the OFAT method, sucrose (X1), inoculum size (X2), and agitation speed (X3) were identified as the variables to be investigated for their capacity to enhance the production of EPS from P. pentosaceus SL4. The optimal levels of these variables were determined by utilizing a Box-Behnken design, a statistical design methodology within the response surface methodology (RSM) framework. This experimental design involved the execution of three replicates, encompassing twelve factorial points and five central points (Table 2). The measurement of the resultant EPS yield served as the response variable in this study. Therefore, a quadratic model was developed for EPS yield as represented below in terms of core values: X 1 is sucrose concentration, X 2 is agitation speed, and X 3 is inoculum size. [Table 2] [Table 3] The triplicate responses from each run were analysed using one-way ANOVA (Table 3). The quadratic polynomial model derived from Response Surface Methodology (RSM) effectively characterized the relationship between sucrose concentration (X₁), agitation speed (X₂), and inoculum size (X₃) on exopolysaccharide (EPS) production (Figure 2). ANOVA results confirmed the statistical significance of the model (F-value = 14.56, p = 0.001), validating the inclusion of linear, interaction, and quadratic terms. Regression coefficients revealed significant negative linear effects of X₁, X₂, and X₃, as well as notable antagonistic interactions between the factors (X₁X₂, X₁X₃, and X₂X₃), suggesting synergistic modulation of EPS synthesis. The quadratic terms were also significant, with inoculum size (X₃²) exhibiting the greatest curvature effect, thereby delineating an optimal response surface. Optimization analyses, supported by three-dimensional response surface plots, identified the optimal fermentation conditions as approximately 45–50 g/L sucrose, 90–100 rpm agitation, and 6% inoculum size, corresponding to maximal EPS yield. These findings highlight the critical influence of both individual and combined fermentation parameters and emphasize the necessity of multifactorial optimization in maximizing EPS biosynthesis efficiency. The optimal value, predicted using the BBD experimental model, was selected for the verification test. The verification test involved four repeated experiments by setting sucrose, agitation speed, and inoculum size at predicted levels: 50 g/L, 100 rpm, and 6%, respectively. The results showed no significant difference (p>0.05) between the experimental and predicted values, suggesting that the model can predict the EPS yield. [Figure 2] The molecular weight of EPS The molecular weight of EPS was estimated by the polyethylene glycol (PEG) as standard, which has a known molecular weight. Gel permeation chromatography was used to fractionate the crude EPS into two components eluted by deionized water. The polysaccharide content was detected at 94.81% (EPS-1) and 5.19% (EPS-2), indicating that the production of EPS from P. pentosaceus SL4 after purification was approximately 405.79 mg/L (peak 1) and 22.21 mg/L (peak 2). The presence of sharp, single, and symmetrical peaks in gel permeation chromatography suggests that EPS is a homogeneous fraction. The molecular weight of the exopolysaccharide (EPS) was estimated to be 7.35×10 5 KDa for EPS-1 and 2,7 KDa for EPS-2. Functional group analysis of EPS FT-IR spectroscopy has been used to investigate the presence of functional groups qualitatively and the linkage among the monosaccharides of the EPS. The absorbance was recorded using FT-IR at a wavenumber from 4000 to 500 cm -1 . The IR spectrum of the EPS from P. pentosaceus SL4 revealed absorption characteristics for polysaccharides, which had many peaks from 3406 to 581.19 cm -1 . A broad peak around 3406 cm -1 corresponds to O–H stretching vibrations, which are typical of polysaccharides due to the abundance of hydroxyl groups in sugar monomers. The peaks at approximately 2920 and 2850 cm⁻¹ are attributed to C–H stretching vibrations of aliphatic –CH₂ and –CH₃ groups, indicating the presence of alkyl chains. A distinct peak at 1653 cm -1 is attributed to the C=O stretch bond vibration. In addition, the weak absorption peak observed at 1542 cm -1 is related to the N-H. A weak absorption at 1417 cm -1 is related to the presence of C-H, while intense absorption peaks at around 1219 and 1053 cm -1 are assigned to carboxylic acid and ester groups C-O stretching vibrations. Several minor peaks associated with α- or β-glycosidic configurations and ring vibrations of polysaccharides were also observed in the absorbance range of 914–581 cm⁻¹. Overall, the spectral features confirm the presence of hydroxyl, carbonyl, and glycosidic functional groups, supporting the identification of the sample as a complex exopolysaccharide. [Figure 3] Microscopic manifestation of EPS Scanning electron microscopy (SEM) is a technique for illustrating common physical properties, including microstructure, surface morphology, and texture of EPS. As illustrated in Figure 4, the SEM of EPS produced by P. pentosaceus under optimal conditions (sucrose concentration of 50 g/L, agitation speed of 100 rpm, and inoculum size of 6%) with incubation at 37°C for 24 h displays a compact and porous structure with a flake-like morphology, as well as irregular, overlapping aggregates of varying size. The Energy Dispersive X-ray (EDX) has become a staple quantitative elemental sample analysis method. The EDX analysis revealed that carbon and oxygen were the predominant elements, with a mass ratio of 48.33% and 42.72%, respectively, suggesting EPS is comprised mainly of carbohydrates. Heteroatoms such as phosphorus (P) and nitrogen (N) were detected at approximately 0.29% and 7.91%, respectively. In addition, the analysis revealed traces of sulphur (0.21%) in the EPS. [Figure 4] Thermal properties of EPS The thermal properties of the EPS were found to be critically important for the intended application. Differential Scanning Calorimetry (DSC) was employed to analyse the thermal behaviour of the sample, including melting temperature, glass transition temperature, and crystallization characteristics. The thermal characterization of the EPS revealed the presence of both endothermic and exothermic transitions, observed within the temperature range of 30–400 °C for the EPS derived from Pediococcus pentosaceus SL-4. The DSC thermogram of the EPS exhibited a broad endothermic peak, beginning at 46.8 °C and ending at 127.8 °C, with a melting point peak observed at 91.7 °C. The enthalpy of fusion (ΔH) was calculated to be 232.2 J/g. A second endothermic peak was detected at 226.1 °C, with an associated enthalpy change of 29.21 J/g, corresponding to partial melting or the onset of thermal degradation. No detectable glass transition temperature was observed in the DSC thermogram of the EPS sample. Finally, a mild exothermic drift beyond 250 °C was observed, suggesting the initiation of degradation of the polymer backbone.Top of Form [Figure 5] Technological Properties Microbial polysaccharides have been identified as a promising renewable source for hydrocolloids in the food industry, owing to their distinctive functional characteristics, including high solubility and substantial water-retention capabilities, and act as stabilizer. In this study, the findings indicate that the water solubility index and water holding capacity of EPS were 21.53 ± 0.58% and 130.52 ± 1.44%, respectively. Thereby to preventing disintegration into their native phase and thus enhancing the stability of food products, in this study the probable effect of EPS concentration on suspension stability against virgin red palm oil was investigated. Emulsification activity (EA) values were tested by adjusting EPS at several concentrations (0.2-1.0%). A statistically significant difference (p<0.05) in the EA values was observed when the emulsion was prepared with concentrations ranging from 0.2 to 1% (Figure 6). Therefore, EPS at 0.6% is the optimal concentration for EPS when used as a stabilizer. No significant difference in its emulsifying capacity is observed at higher concentrations. The stability of the emulsion was determined by allowing it to sit for a more extended period, up to 72 hours, after which the emulsion gradually decreased after 24 hours. A common challenge in applying emulsifiers is maintaining the emulsion formed under extreme conditions, such as high and low temperatures and varying pH levels. This study evaluated the emulsion-forming and stabilizing properties at varying temperatures (0, 30, 60, and 90°C) and pH levels (3-7). The emulsion of EPS with virgin red palm oil exhibited stability within the pH range of 4-7 and emulsification stabilizer declined at storage temperature above 30°C. [Figure 6] 4. Discussion Sourdough prepared using tomi-tomi fruit ( Flacourtia inermis ) starter is a source of various lactic acid bacteria species (Kemhay et al. 2024 ). Pediococcus pentosaceus SL4, one of the lactic acid bacteria successfully isolated, has the potential to produce a considerable number of exopolysaccharides. In the screening process using MRS Agar medium supplemented with 0.1% Congo Red indicator, P. pentosaceus SL4 demonstrated the ability to produce exopolysaccharides, as indicated by a color change of the MRS Agar medium to dark brown. Several studies have also reported similar findings, where P. pentosaceus was successfully isolated from sourdough made with grape, apple, and pomegranate starters (Gordún et al. 2015 ; Ripari et al. 2016 ). The exopolysaccharides produced by certain lactic acid bacteria during dough fermentation have also been reported to enhance the sensory characteristics of sourdough (Zhang et al. 2023b ). Other research suggests that exopolysaccharides hold potential as functional food ingredients or additives to improve the physical and chemical properties of food products (Loeffler et al. 2020 ). Those potentials can be further explored by enhancing P. pentosaceus SL4 to produce EPS then characterize them. Many factors contribute to the enhanced production of exopolysaccharides (EPS) by lactic acid bacteria. The presence of an excess carbon source in the form of saccharides in the fermentation medium can enhance the ability of lactic acid bacteria to produce exopolysaccharides. This phenomenon is associated with an increase in osmotic pressure due to the high sugar content. Several studies have shown that the maximum EPS yield (53.79 g/L) was attained with Leuconostoc mesenteroides DRP105, utilizing sucrose at 86.83 g/L as a carbon source (Xing et al. 2018 ). A parallel finding was reported for Leuconostoc citreum B-2, which yielded 59.33 g/L with sucrose supplementation at 99.8 g/L (Wang et al. 2020 ). P. acidilactici MT41-11 was reported to be able to produce exopolysaccharides of 500 mg/L when fermented MRS media by adding 5% sucrose (Bai et al. 2021 ). However, other studies have acknowledged that alternative carbon sources, such as maltose and lactose, can stimulate EPS production by Enterococcus and Lactobacillus (Imran et al. 2016 ; Zanzan et al. 2023 ). The variation in EPS production can be attributed to differences in bacterial strains and the type of carbon source utilized (Imran et al. 2016 ). Each strain has a different metabolic pathway, so it uses different carbon sources. High sucrose concentrations in media induced the β-fructofuranosidase enzyme production from microorganisms to catalyse the conversion of sucrose into glucose and fructose, which can later be used as substrates (Sheng et al., 2016 ). In addition to the critical role of carbon source selection and concentration, agitation as a parameter of the fermentation process has been identified as a factor that also significantly affects EPS production. Agitation also plays a role in maintaining the homogeneity of the medium throughout the fermentation process. The inadequate mixing of high-viscosity media gives rise to non-homogeneity. Nevertheless, elevated mixing speed engenders elevated shear stress in the culture medium, diminishing the EPS yield (Prasad and Purohit 2023 ). Fermentation of L. rhamnosus in a mixed medium of fructose, glucose, and saccharose at 20 g/L with an agitation speed of 80 rpm, which exhibited augmented EPS production in comparison to fermentation conducted without agitation (Oleksy-Sobczak et al., 2020 ). Furthermore, previous studies have also investigated the optimization of exopolysaccharide production through the regulation of inoculum concentration; however, inoculum concentrations exceeding 10% did not result in a significant improvement in EPS yield from an economic standpoint (Oleksy-Sobczak et al., 2020 ). Beyond agitation and inoculum concentration, fermentation time emerges as a critical parameter affecting EPS yield. Precise determination of the optimal fermentation duration for each microorganism is vital, as mismatches between the growth and EPS production phases can compromise both efficiency and yield. Extended fermentation periods often result in decreased EPS production, largely due to enzymatic degradation by glycosyl hydrolases (Haj-Mustafa et al. 2015 ; Amiri et al. 2019 ), including rhamnosidase produced by species such as Lactobacillus plantarum (Ferreira-Lazarte et al. 2021 ) and Lactobacillus acidophilus (Mueller et al. 2018 ). This pattern aligns with observations from Haj-Mustafa et al. ( 2015 ), who noted peak EPS synthesis by L. rhamnosus 519 and L. acidophilus LA5 within the first 49 and 24 hours, respectively, followed by gradual declines. Similarly, Enterococcus mundtii A2 demonstrated maximum EPS yield at 16 hours, with reduced production at longer incubation times (Zanzan et al. 2023 ). In the RSM optimization, by adjusting sucrose concentration, agitation speed, and inoculum concentration to optimal conditions, the EPS yield reached 428.0 mg/L, which is 7.8-fold higher than the original yield (55 mg/L in MRS medium, Table 1 ). Moreover, this yield was also higher than those reported for Enterococcus mundtii A2 (382.27 mg/L) (Zanzan et al. 2023 ), Lactobacillus plantarum NTMI05 (197 mg/L), and Lactobacillus plantarum NTMI20 (187 mg/L) (Imran et al. 2016 ). However, some other species such as Leuconostoc lactis L2 (58.1 g/L) (Jiang et al. 2020 ), Lactobacillus plantarum KX041 (599.52 mg/L) (Wang et al. 2017 ), and Enterococcus faecium MC-5 (16.48 g/L) (Tilwani et al. 2021 ) have been reported to produce even higher amounts of EPS. In addition to strain differences, variations in fermentation optimization parameters can also influence the EPS yield obtained. The molecular weight of the first fraction of EPS is higher than that of other EPS produced by coccus genera of lactic acid bacteria, which range from 2.5×10 1 KDa to 2×10 3 KDa (Jia et al. 2019 ; Bai et al. 2021 ; Zhou et al. 2021 ; Jiang et al. 2022 ). However, it is nearly equivalent to the molecular weight of dextran, which is produced by fructophilic lactic acid bacteria, Apilactobacillus waqarii strain HBW1 (Ahmad et al. 2022 ). These discrepancies in molecular weight may be attributable to inter-species and inter-strain variations. The elevated molecular weight of EPS produced by lactic acid bacteria renders them a viable alternative to stabilizers in the food industry, biomedicine, and pharmaceuticals. The FTIR spectrum of the EPS in this study aligns with previous findings. A broad band at 3590–3389 cm⁻¹ corresponds to O–H stretching, indicating abundant hydroxyl groups typical of polysaccharides (Amiri et al. 2019 ; Zanzan et al. 2023 ). This region is also associated with water solubility (Vinothini et al. 2019 ). The peak at around 2932 cm⁻¹, attributed to C–H and C–O stretching, matches that reported in EPS from L. plantarum KX041 (Wang et al. 2017 ). A peak at 1633 cm⁻¹ corresponds to C = O stretching, as also found in EPS from L. acidophilus LA5 (Imran et al. 2016 ). A weak absorption peak at 1542 cm⁻¹ is related to N–H bending of amide II from proteins (Wang et al. 2017 ). Notably, absorption in this region was not observed in EPS from other P. pentosaceus strains (Jiang et al. 2022 ). The fingerprint region (800–1200 cm⁻¹) shows typical polysaccharide features, with 1200–1000 cm⁻¹ indicating C–O and C–O–C glycosidic linkages (Amiri et al. 2019 ). A band at 813.44 cm⁻¹ suggests α-glycosidic linkages (Zanzan et al. 2023 ). These features confirm that the EPS contains characteristic functional groups commonly found in microbial exopolysaccharides. Microscopic analysis revealed that the morphology of exopolysaccharides (EPS) produced by different bacterial strains can vary significantly, even among those belonging to the cocci group. For example, levan produced by Enterococcus faecium MC-5 exhibits a uniform spherical morphology (Tilwani et al. 2021 ), whereas glucan synthesized by Enterococcus hirae KX577639 presents an irregular and interconnected structure (Jayamanohar et al. 2018 ). Such variations in EPS microstructure are primarily attributed to differences in the composition of monosaccharide units, chain length, and the types of glycosidic linkages forming the polymer backbone. Furthermore, EPS characterized by porous and spongy textures, as observed in this study, possess enhanced water-binding capabilities, which may contribute to their functional properties. This property that lends itself to applications in the food industry as a gelling, thickening, and stabilizing agent. This characteristic renders it a promising candidate for producing edible films and coatings for food products (Jiang et al. 2022 ). The heteroatom P and N in the partially purified EPS P. pentosaceus SL4 proposed a trace of phospholipids and protein, respectively (Zaghloul and Ibrahim 2022 ; Zanzan et al. 2023 ). Also, the analysis revealed traces of another element, including sulphur, which were close to the EPS produced by Leu. mesenteroides DRP105 (Xing et al. 2018 )d sakei L3 (Wang et al. 2019 ). The elemental composition of the SL4-EPS was similar to the EPS produced by Enterococcus sp.BE11 (Zaghloul et al. 2023 ) and Enterococcus mundtii A2 (Zanzan et al. 2023 ). The DSC analysis of the exopolysaccharide (EPS) sample demonstrated thermal behaviour consistent with previous studies on similar biopolymers. This analysed was employed to ascertain the correlation between physicochemical properties and temperature during application (Vinothini et al. 2019 ). The prominent endothermic peak, attributed to the loss of bound water, which is facilitated by the abundant carboxyl groups in EPS that bind with water molecules. As the temperature rises, the hydrogen bonds that bind the water molecules weaken, releasing them (Tilwani et al. 2021 ). This is in line with the study that has been conducted by Jiang et al. ( 2022 ), who observed dehydration peaks in EPS from P. pentosaceus E-8 occurring at 82.6°C due to the hygroscopic nature of polysaccharides. The relatively high enthalpy value in first endothermic peak in this study suggests a significant water-binding capacity, which is critical for the material’s physical properties and stability. The second endothermic phase occurred between 226 and 233.8°C, corresponding to the higher protein content in the EPS. Higher temperatures associated with higher energy requirements are necessary to remove proteins covalently bound to polysaccharides (Wang et al. 2017 ). Studies on microbial EPS have demonstrated thermal stability up to approximately 220–260°C before degradation initiates, consistent with the mild exothermic transition observed here beyond 250°C. For instance, EPSF2 from Enterococcus sp. F2 maintained structural integrity up to 266.6°C (Jiang et al. 2021 ) and EPSE8 from P. pentosaceus exhibited stability up to 257°C in TGA analysis (Jiang et al. 2022 ). The slight variation in peak temperature and enthalpy values may be attributed to differences in molecular weight distribution, degree of branching, and molecule structure (Lakra et al. 2020 ; Jiang et al. 2021 ). Solubility, a pivotal metric in assessing the saturation concentration of EPS in an aqueous environment at a designated temperature, is a critical factor in determining the textural characteristics of food products (Jiang et al. 2022 ). Water-holding capacity is the ability to hold an immense amount of water (Vinothini et al. 2019 ). The water solubility of EPS in this study was nearly higher than that of EPS extracted from Leu. lactic KC117496 (14.2%), yet the water-holding capacity value obtained was lower (Saravanan and Shetty 2016 ). In contrast, D-glucan produced by Enterococcus hirae KX577639 exhibited a water solubility index of 46.5% and a water holding capacity of 202.04%, which is higher than the results obtained in this study (Jayamanohar et al. 2018 ). The molecular weight and percentage of branching appear to influence their water solubility (Jayamanohar et al. 2018 ). Furthermore, the high water-holding capacity of EPS can be attributed to its porous polymer structure, which enables water retention through hydrogen bonds (Saravanan and Shetty 2016 ). The elevated WHC of EPS indicates its capacity to retain substantial amounts of water, thereby mitigating the dehydration levels of the product. A previous study has demonstrated that incorporating 4% EPS into minced meat can stabilize the meat during cooking, thereby reducing losses (Wu et al. 2024 ). EPS's high solubility and water-retention capacity render it a suitable stabilizer and bio-thickener. Xanthan gum polymer has been reported to have a solubility index of 64.86% and a water holding capacity of 1000.33% (Sarkar et al. 2018 ), respectively, and has been recommended by the US as a stabilizer, thickener, binder, and texturizer. EPS from P. pentosaceus SL4 has been identified as a potential alternative renewable hydrocolloid despite xanthan gum's higher solubility and water binding. This finding is consistent with the observations reported for EPS derived from L. plantarum ATCC 8014, which exhibited emulsification activity of approximately 60% at 72 hours (Balyan et al. , 2024). However, a separate study demonstrated that the emulsifying properties of a mixture of lactic acid bacteria found in yogurt could stabilize limonene emulsions by over 70% after 72 hours (Balyan et al. 2024b ). Notably, the optimal emulsifying capacity of EPS produced by P. pentosaceus SL4 is about 74.75% when employed in products with a temperature of 4°C and a pH range of 4–7. Furthermore, a comprehensive study established that a reliable emulsifier must retain at least 50% of the original emulsion volume after 24 hours of its formation (Vinothini et al. 2019 ). Declarations Conflict of Interest The authors declare no conflicts of interest related to the results of this study with other parties Author Contribution DAP: study design, data acquisition, data analysis and interpretation, drafting the manuscript; RY: material support, critical revision of the manuscript; CH: study design, study supervision, critical revision of the manuscript; DAS: study concept and design, study supervision, data interpretation, critical revision of the manuscript. Acknowledgement This study was supported by the Indonesia Endowment Fund for Education Agency (LPDP) and the National Research and Innovation Agency (BRIN). 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Int J Biol Macromol 192:716–727. https://doi.org/10.1016/j.ijbiomac.2021.10.047 Tables Table 1 Effect of carbon source on cell growth and EPS produced by P. pentosaceus SL4 Carbon Source Saccharide Cons (g/L) Growth (OD 600 nm) EPS yield (mg L -1 ) MRS+Glucose 20 0.138 a 75.0±0.00 a MRS+Sucrose 20 0.154 a 85.0±10.61 a MRS+ Lactose 20 0.141 a 60.0±7.07 a MRS+ Fructose 20 0.133 a 60.0±0.00 a MRS+ Galactose 20 0.149 a 76.0±17.67 a MRS - 0.139 a 55.0±5.67 a a Values in the same column not followed by a common superscript letter are significantly different (p < 0.05) Table 2 BBD matrix along with actual and predicted EPS yield (n=3) Variables Response EPS (mg/L) Run Sucrose (g/L) Agitation (rpm) Inoculum (%) Actual Predicted 1 50 150 4 344,9 332,6 2 50 50 4 346,2 351,2 3 50 50 8 347,5 359,8 4 75 100 8 315,1 306,5 5 25 150 6 388,8 392,6 6 75 150 6 305.3 318,9 7 50 100 6 463.3 440,2 8 75 100 4 359 357,7 9 25 50 6 412,3 398,7 10 50 100 6 396,5 418,1 11 25 100 4 350,3 359,1 12 50 150 8 295,7 290,7 13 25 100 8 375,7 377,0 14 75 50 6 404,4 400,6 15 50 100 6 426,2 418,1 16 50 100 6 428 418,1 17 50 100 6 419,8 418,1 Table 3 . Analysis of variance for quadratic model of exopolysaccharide production Source Sum of squares df Mean square F-value p-value Model 28832.90 9 3203.66 14.56 0.0010 significant A-Sucrose_Conc 2574.03 1 2574.03 11.70 0.0111 B-Agitation 3858.81 1 3858.81 17.53 0.0041 C-Inoculum_Conc 554.44 1 554.44 2.52 0.1565 AB 1428.84 1 1428.84 6.49 0.0382 AC 1193.70 1 1193.70 5.42 0.0527 BC 637.56 1 637.56 2.90 0.1325 A² 602.28 1 602.28 2.74 0.1420 B² 3410.41 1 3410.41 15.50 0.0056 C² 13244.33 1 13244.33 60.18 0.0001 Residual 1540.49 7 220.07 Lack of Fit 903.42 3 301.14 1.89 0.2723 not significant Pure Error 637.07 4 159.27 Cor Total 30373.39 16 Fit Statistics Std. Dev. 14.83 R² 0.9493 Mean 372.71 Adjusted R² 0.8841 C.V. % 3.98 Predicted R² 0.4913 Adeq Precision 11.2024 Additional Declarations No competing interests reported. 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line) and yield of EPS (bar)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/e7b12518df4fe2fcab35e49c.png"},{"id":91524830,"identity":"4b37f392-aec4-43d3-8214-4bf20545d220","added_by":"auto","created_at":"2025-09-17 11:02:43","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":459169,"visible":true,"origin":"","legend":"\u003cp\u003eThe surface plots of EPS production by \u003cem\u003eP. pentosaceus\u003c/em\u003evs. (a) agitation, sucrose (inoculum size and fermentation time: 6% and 24 h, respectively), (b) inoculum, sucrose (agitation and fermentation time: 100 and 24 h, respectively), (c) inoculum, agitation (sucrose concentration and fermentation time: 50 g/L and 24 h, respectively)\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/9261c3b1724455a937042c41.png"},{"id":91524832,"identity":"3669b189-5fe4-417c-9cf9-8601f43e3429","added_by":"auto","created_at":"2025-09-17 11:02:43","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":156360,"visible":true,"origin":"","legend":"\u003cp\u003eElution profile of crude EPS on gel chromatography column at a flow rate of 0.3 mL/min and FT-IR spectrum of EPS produced by \u003cem\u003eP. pentosaceus\u003c/em\u003e SL4\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/9a5d4760f5d75a8642a2c546.png"},{"id":91526647,"identity":"13b6afc2-1ef7-435a-b509-c73bfcff5253","added_by":"auto","created_at":"2025-09-17 11:10:43","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":474919,"visible":true,"origin":"","legend":"\u003cp\u003eScanning electron micrograph of EPS at different magnifications: a, 250 ×; b, 2000 ×; c, 15,000 × and the elemental composition of EPS produced by \u003cem\u003eP. pentosaceus \u003c/em\u003eSL4\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/36a9838ef7d55e8317b2ffb8.png"},{"id":91524837,"identity":"f2185138-7d35-4993-bd69-aa99b23f6ab6","added_by":"auto","created_at":"2025-09-17 11:02:43","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":132774,"visible":true,"origin":"","legend":"\u003cp\u003eDSC thermogram EPS from \u003cem\u003eP. pentosaceus \u003c/em\u003eSL4\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/d37c629d6f199ea52f64a3d9.png"},{"id":91524836,"identity":"779b3614-c9a7-4bcf-8a7e-2c94fffe84c3","added_by":"auto","created_at":"2025-09-17 11:02:43","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":72185,"visible":true,"origin":"","legend":"\u003cp\u003eEmulsification activity of emulsion prepared with virgin red palm oil at a) different concentrations, influenced by b) temperature and c) pH. The statistic differences were indicated with different lowercase letters (p\u0026lt;0.05)\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/3668e370f997cdf0263ac0c3.png"},{"id":105755081,"identity":"acc3c6f6-b6fd-4f11-9c8b-ec56df5275c8","added_by":"auto","created_at":"2026-03-30 16:25:04","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2417295,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/4697c376-4045-4d00-b5c0-dd7d370dd8bd.pdf"},{"id":91524834,"identity":"1b74b1fe-14b0-4f28-bd56-fe097c9f0622","added_by":"auto","created_at":"2025-09-17 11:02:43","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":361244,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical Abstract\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"GA.png","url":"https://assets-eu.researchsquare.com/files/rs-7555681/v1/2cd7ec85c3574b6bc3b36714.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Optimization on production and technological properties of exopolysaccharide from Pediococcus pentosaceus SL4 isolated from sourdough Tomi-tomi (Flacourtia Inermis Roxb)","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eThe demand for biomolecules that offer technological and health benefits has been increasing and must be addressed by exploring alternative biomolecule sources with specific properties. Exopolysaccharides (EPS) are a type of extracellular polysaccharide produced by bacteria that have wide applications in the food, cosmetic and pharmaceutical industries, functioning as a stabilizer, texturizer, emulsifier, and gelling agent (Wang et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Balyan et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024a\u003c/span\u003e). Some groups of lactic acid bacteria (LAB), such as \u003cem\u003eLactobacillus\u003c/em\u003e, \u003cem\u003eEnterococcus\u003c/em\u003e, and \u003cem\u003ePediococcus\u003c/em\u003e, produce EPS through sucrose supplementation in the fermentation medium (Jayamanohar et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Wang et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Jiang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) to protect themselves from environmental stresses such as insufficient nutrients, pH, salinity, temperature, or co-cultivation (Wang et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Vinothini et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Oleksy-Sobczak and Klewicka \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Jiang et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Depending on the sugar composition, EPS can be either homopolysaccharides or heteropolysaccharides. The properties of EPS are associated with their characteristics, such as molecular weight, thermal properties, and sugar composition. EPS produced by LAB has the potential to play an important role because it has been classified as Generally Recognized as Safe (GRAS). The properties of EPS vary depending on the strain and fermentation conditions. Consequently, screening for LAB shows potential EPS production is necessary. Fermented foodstuffs like sourdough could be an ideal source for EPS-producing LAB. LABs exist spontaneously in sourdough due to the materials utilized or the environment where the sourdough is produced (Ripari et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Yu et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Some LABs, including \u003cem\u003eP. pentosaceus\u003c/em\u003e, \u003cem\u003eL. plantarum\u003c/em\u003e, and \u003cem\u003eL. brevis\u003c/em\u003e, have been isolated from sourdough and produce EPS (Ripari et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Fu et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The presence of exopolysaccharides produced by LAB has been shown to improve the rheological properties of sourdough (Zhang et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2023a\u003c/span\u003e) and enhance health benefits such as antioxidants (Wang et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), maintain blood sugar stability, and reduce cholesterol (Dilna et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Other studies reported that the utilization of lactic acid bacteria, such as \u003cem\u003eLactobacillus spp., Pediococcus spp., and Leuconostoc spp\u003c/em\u003e., in the production of EPS has been demonstrated to enhance the stability and quality of food products through its application as a stabilizer and a coating agent (Wang et al, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Jiang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Balyan et al. 2024).\u003c/p\u003e\u003cp\u003eWe developed the sourdough using some tropical fruits as a starter (Kemhay et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) and we also isolated \u003cem\u003ePediococccus pentosaceus\u003c/em\u003e SL4 were among the LAB species and had been screened for its potential EPS production (unpublished data). \u003cem\u003eP. pentosaceus\u003c/em\u003e cannot produce large amounts of exopolysaccharides without optimal conditions, while under optimal conditions, LAB produces EPS that can be 2 to 5 times higher depending on the bacterial strain (Xing et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Wang et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Jiang et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). The optimization of carbon sources and fermentation process conditions has been identified as a strategy to enhance EPS production (Oleksy-Sobczak et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). However, it is crucial to note that each strain exhibits distinct optimal conditions for EPS production. The objectives of this study are to optimize EPS production of \u003cem\u003ePediococccus pentosaceus\u003c/em\u003e SL4 by adjusting carbon source, agitation speed, inoculum size, and fermentation time and subsequently derive a quadratic model to biosynthesize EPS and to characterize the EPS using various instrumental analyses and estimate its technological properties as a stabilizer in food processing.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eMaterials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe high-yield EPS-producing strain \u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 was isolated from \u003cem\u003etomi-tomi\u003c/em\u003e sourdough and was stored at -80⁰C in the Biotechnology Laboratory Faculty of Agricultural Technology Gadjah Mada University. de Man Rogosa Sharpe (MRS) broth (Himedia, India) supplemented with saccharides was used to cultivate the SL4 for EPS production. Glucose, fructose, galactose, and lactose as supplemented sugar were from Merck (Darmstadt, Germany) and sucrose from Phytotech (Kansas, USA). All chemicals used for analysis were analytic grade from Sigma Aldrich (Missouri, USA), and virgin red palm oil was from the local Indonesian market.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreliminary screening of fermentation variables and optimization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eScreening design is a methodical approach that facilitates the comprehension of the factors potentially implicated in the system. The bacteria were inoculated in MRS broth for 18 hours at 37\u0026deg;C for pre-culture. The first stage in screening was determining the right carbon source. Five types of carbon sources at 20 g/L were supplemented to MRS broth, culture at concentration 2% (v/v) was inoculated into sterile broth, and fermentation was carried out at 37⁰C with 80 rpm agitation for 48 hours. The best carbon source screening results were used for the next screening stage. In the second stage, the variables utilized were sucrose (25\u0026ndash;125 g/L), inoculum size (2\u0026ndash;10%), agitation speeds (0-200 rpm), and cultivation time (8\u0026ndash;56 h) used to evaluate the maximum EPS production. The starter was inoculated at specific conditions based on one factor at a time (OFAT) with four numerical factors. After identifying the factors affecting EPS production by OFAT design, a Box-Behnken design (BBD) was employed to assess the impact of three additional factors. The 2% culture (v/v) was inoculated to MRS broth (5% glucose) and incubated at 80 rpm at 37⁰C for 48 h, which was used as a non-optimized fermentation condition.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExtraction and purification of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe extraction of exopolysaccharide (EPS) from \u003cem\u003eP. pentosaceus\u003c/em\u003e was performed according to the method of Zanzan et al. (\u003cspan class=\"CitationRef\"\u003e2023\u003c/span\u003e) with modifications. After incubation, the cultures were heated at 100\u0026deg;C for 10 minutes to inactivate EPS degrading enzymes, followed by centrifugation at 5000\u0026times;g for 30 minutes. The pre-chilled 96% (v/v) ethanol was added to the resultant precipitate twice the initial volume. The mixture was then stored at 4\u0026deg;C for 24 hours, followed by precipitation using centrifugation at 5000\u0026times;g for 30 minutes at a temperature of 4\u0026deg;C. Subsequently, 8% (w/v) trichloroacetic acid (TCA) was added to the precipitates and maintained at 4\u0026deg;C for 30 minutes to yield a protein precipitate. The protein precipitates were then removed by centrifugation at 5000 \u0026times; g for 30 minutes. Subsequently, two volumes of pre-chilled 96% (v/v) ethanol were added to the filtrate and stored at 4\u0026deg;C for 24 hours, and the precipitate was subjected to centrifugation at 5000\u0026times;g for 30 minutes at 4\u0026deg;C. The polysaccharide pellet was dissolved in 5 mL of deionized water and dialyzed using a 12\u0026ndash;14 kDa cut-off dialysis membrane. The resulting EPS was then lyophilized, resulting in a powder form of the EPS suitable for further analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCharacterization of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMolecular weight analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSize exclusion chromatography was used to determine the molecular weight distribution of EPS. Ten milligrams of EPS sample were dissolved in 1 mL of ddH\u003csub\u003e2\u003c/sub\u003eO and filtered through a 0.22 \u0026micro;m filter. Then, the EPS solution (10 mg/mL, 20\u0026micro;L) was loaded into the gel permeation chromatography system, TOSOH HLC-8320GPC Japan, with TSKGel Super AW5000 as column and coupled to the Refractive Index detector. Isocratic elution was carried out using deionized water at a constant rate of 0.3 mL/min, and the detector temperature was set to 40⁰C.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFourier transform infrared (FT-IR) analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eTen milligrams (10 mg) of the lyophilized EPS sample were ground with KBr powder. The spectrum was recorded using a Bruker vertex 70 DTGS FTIR spectrometer (Bruker, Germany) with a resolution of 4 cm\u003csup\u003e-1\u003c/sup\u003e in the 400\u0026ndash;4000 cm\u003csup\u003e-1\u003c/sup\u003e range.\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eScanning Electron Microscopy (SEM) and Energy-dispersive X-ray analysis of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eThe morphology of lyophilized EPS was observed under SEM JEOL JSM-IT200 (Tokyo, Japan). In brief, 5 mg of lyophilized EPS was placed on SEM stubs, tapped with double-sided, and coated with a carbon layer of approximately 10 nm thickness. The image has been taken at 10 kV at a resolution of 250\u0026times;, 2000\u0026times;, and 15000\u0026times; magnification. The same samples for SEM were used for EDX analysis but with an acceleration voltage of 20 kV and at 250x magnification.\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eThermal properties\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDifferential scanning calorimetry (NETZSCH DSC 300 Caliris Select, Germany) was used to investigate the thermal properties of EPS. The EPS sample (3\u0026ndash;5 mg) was put in an alumina pan and sealed. The system was heated from 25⁰C to 400⁰C at a linear heating rate of 10⁰C/min in a nitrogen atmosphere (50 mL/min). The empty aluminium crucible was treated as a reference.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eWater Holding Capacity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWater holding capacity (WHC) was determined by dissolving 10 mg of EPS in 1 mL of double distilled water and mixing in the vortex for 5 min. The sample was then centrifuged at 16000 rpm for 25 min. The pellet was collected and weighed (Marimuthu and Rajendran \u003cspan class=\"CitationRef\"\u003e2023\u003c/span\u003e). The WHC was determined using Eq.\u0026nbsp;(1)\u003c/p\u003e\n\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n \u003cdiv class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\u003cimg src=\"data:image/png;base64,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\" width=\"746\" height=\"78\"\u003e\u003c/div\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eSolubility Index\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe EPS sample (10 mg) was dissolved in 1 mL double distilled water under constant agitation for 24 h, then centrifuged at 6000 rpm for 20 min. A triple volume of chilled ethanol 96% was added to the supernatant to precipitate EPS. It was then centrifuged at 12000 rpm for 20 min. The pellet was dried at 55⁰C and weighed (Marimuthu and Rajendran \u003cspan class=\"CitationRef\"\u003e2023\u003c/span\u003e). The solubility was calculated using Eq.\u0026nbsp;(2)\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\" width=\"746\" height=\"77\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEmulsification properties of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe emulsion index was evaluated by suspended EPS at various concentrations (0.25\u0026ndash;1.25 mg/mL) in double distilled water as described previously (Marimuthu and Rajendran \u003cspan class=\"CitationRef\"\u003e2023\u003c/span\u003e) with slight modification. A 2 mL of EPS solution was added to an equal volume of virgin red palm oil, and the mixture was vortexed vigorously for 5 min. The emulsion activity (%EA) was defined as the percent retention of the emulsion after incubation for one hour at room temperature, whereas emulsion stability (%ES) was determined after 24, 48, and 72 h separately. The %EA and %ES calculated according to Eq.\u0026nbsp;(3)\u003c/p\u003e\n\u003cdiv id=\"Equc\" class=\"Equation\"\u003e\n \u003cdiv class=\"mathdisplay\" id=\"FileID_Equc\" name=\"EquationSource\"\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAuoAAABLCAYAAAAmuKLEAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAABtSSURBVHhe7d3daxtX+gfwr3733WakXpVSike9KO1i2IzTJXUKKdSjNYvZhS1SmmUxZEmQEgq5qbtychFIspGyLxC2sR0olJBWY9KSUtZey4Ua1trSNiJooEsvaokQQq9mrDb9A2YvfjqHmaMZvfhVdr4fEHXOHI3mnDMdPxqf80zM8zwPREREREQ0UP5PLSAiIiIiot3HQJ2IiIiIaAAxUCciIiIiGkAM1ImIiIiIBhADdSIiIiKiAcRAnYiIiIhoADFQJyIiIiIaQAzUiYiIiIgGEAN1IiIiIqIBxECdiIiIiGgAMVAnIiIiIhpADNSJiIiIiAYQA3UiIiIiogHEQJ2IiIiIaAAxUCciIiIiGkAM1GlgWJaFWCyGWCyGYrGobt5VuVxOHlulUlE375pisYhkMjmQx9aLQR7zXu2HNhAR0WBioE4DI5PJoFQqqcVbJhaLIZVKqcU9uX79OrLZrFq8q3K5HG7fvo2vvvoKtVpN3bwnbHbMK5XKrgfIm20DERFRFAbqNFCeeeYZtWhgPPfcc2pRR+Iu/HawbRszMzN44403kEgkMDw8DM/zMDo6qlYdeIM85r3aD20gIqLBw0CdHhue52FpaUkt3jaNRkMt2jLffvstAODw4cPqpsfK6OgoPM/D22+/rW4iIiLa8xioE22DRqOBcrmsFm+ZBw8eqEVERES0zzBQp4FVqVSQSqUQi8UQj8dhWZZaBWgFxblcDvF4HLFYDMlkEnNzc3K7bduBBZeqSqWCTCYjt6uvsAWalmXJfSaTSdi2HdhmGIb8d7d9+VmWhZGREVl/ZGQEi4uLgTrT09O4cuUKAODIkSOI9TD3vlsfAQh8bqPRwNzcnGxjKpWCbdtwXRfFYlHuJ5VKwXVduQ/btgPbupV3Yts2MpmMfJ94r+gPy7I67lNtczweRy6Xazte/zi6rht4j9q+frmui7m5OXkeizEV54tt27JcHIPgX8Acj8cD+/QvIo7H45ieng4cpzqW/vM1rK+IiGhAeUQDZHV11QPgGYbhZbNZz3Ecr16ve7quewC8Wq0WqF+v1z1N02Rdz/O8bDbrAfAWFhZkPcdxPACeesrXajUPgHy/4zjy/YVCIVC3UCjIY5udnfU83/Fqmhao63le6Od1ks/nPQBy3/V63TMMwwPglUqlQF1xLKurq4HyML32ked58vOy2azcd6lUku02TVOWi2PIZrOBfSwsLHgAPNM0eyoXfejvb8dxPE3TPMMwvHq97nme583OzrbVi9qnaHPY+w3DkP3g+c4NXde9bDYrz7Go9oUJa0OtVvM0TfN0XZfH4D+HBH//qkzT9DRNk+/3WmPkb5d4fz6f970zOJazs7OyntpXREQ0uHqPIoh2gAh41GBCBDgiiBXS6bSnaVog8PJaQbKu621lUAJnEbD6AyERuEUdgxrAm6bpIeRLRNjnRfF/YfCr1+seQr4I9BOo99NHoi3qFwPxRUn9PIQEmFFj2K3c369hZV7r+MLqqftMp9MelHH1fOOtnkdirNT6Ye0LE3a8okztS03TPCjnha7rbWPstYJtfwAuvmyo4yDGzX/8oszf1nQ63dZXREQ0uDj1hQbSa6+9Fvi3WDT5448/Bsrn5+dx6NAhJBKJQLlpmqjX64GyMGLB59DQkCxT96VSF3CKY/3pp58C5f0Q89lfffXVQPnQ0BBM00Sz2QxMr+nHRvpIzWIipmSEZZWpVqtq0aY9/fTTAIArV64EpjwtLS31tHB0fn4euq4HxhUA3nzzTQDAyspKoFxQ62MT7RMLXTOZTKD80KFDgX8DwMmTJ9FsNgPTnGzbRrVaxYkTJ2TZnTt3gJBxEOfg999/HygHgBdffFH+bFnWji6oJiKizWGgTnuWmO9dLpcD83xjsZgMfLvNCReBmT9Di5jre+DAAVm23T7//HMgJED228gXga3oo90wNDQkc5MfO3YMyWSy51zpoj3++d6qH374QS3aVb/73e8AADdv3pRlc3NzyGazgS8PYszUsZyamgIAfPHFF7IuERHtfQzUac8rFApoTeNqe6l3HlXiDms+n4frunBdF+fPnwcAnDlzRqm9d22mj3ZLJpPBd999h9nZWQDA1NQURkZGNrW4c6eJxaSZTEYu8AzLBiT+cjI/Py/Pw5mZGZw6dUqtCtM028ZQvHr5awMREe0dDNRpz3rhhRcAAPfu3VM39Wx0dBT5fB7Ly8t46qmn8NRTT+Hu3btYWFjYdACraZpaFOngwYMAgIcPH6qbpCeeeEIt6mor+qhfYtrKVkgkEjh16hTW1taQzWZRrVbx0UcfqdUCRJvX1tbUTdJO/LXEtm08//zzWFlZwZ/+9CfcvXsXnufBNE21KgBgcnISAPDZZ5/hvffeg2maGB4eDtQxDANff/11oIyIiPYvBuq0ZyUSCRiGgeXl5Q3fZV1cXMT8/Dy+++47eVfy7t27GB8fV6v2LWwucpRXXnkFAPD+++8Hyl3XRblchq7rbUFbL7aij/oVNs8bAB49eqQWRSoWi8jlcoEy8dcPdZ2CSrS5Xq+3Tev517/+BQD4zW9+EyjfDn/+85/RbDbx7rvvBsYu6gtEJpOBpmn461//ihs3buDcuXNqFYyNjaHZbLa1i4iI9icG6jRQxB3l+/fvB8rF3Fs1SLtw4QKazSZOnz4t55k3Gg0Ui0VMT0/Lev4g1f/ztWvX5M/diGNS73qL+eVqICoCVpGvfG5uLjIX/Pj4OEzTRLlclvVd18Xx48eBkABe9IMIPDvptY/g6xu1LSK4DJvLr/4MALquo1wuy4CyUqngn//8Z2jdqDGfmZkJ9MWHH34ItKZ+COI41X3+/e9/BwCcPXtWHvPi4iIuX74M0zQDCzyj2hH1c5iwNojc5+IvACKvu/hLS9iTazOZjFy8GvYXnRMnTkDTNJw9e1b2reu6sCyrLT961FgSEdEeoqaBIdotIs+zeIlUhWq5mlpvYWFB5owW6fQKhUIgHaFILwgl1aFIixj2SqfTMuWiSOsnXiLlnlruT9HoOI5MkafreluaPpXjOF6hUAgcq/8YBJGiT7zU/gjTSx9FtUWkOkSr70S5SDMoyv1qtZr8PF3XZdpCUV+khVTHVoz56uqqzCEutpmmGcj7LvKUi+3pdFpuE/vwH7v/OPyizo1O7fOLaoPjOIHPT6fTXr1el6k1/X0piPOx07lSq9Xa2pXNZgOpGdWx7LQ/IiIaXDHv/395Ej2WUqkUDhw40Hanu1KpYGJiAmNjY23biLZLpVLB5ORk5PQYIiJ6vHDqCz22KpUKyuVyaHaX0dFR6Lo+cGn8aH+pVCoYGRkBWlNVJicncfHiRbUaERHtsEqlgkwm03NqYD/LspDJZLZkPdGmA3XbtpFMJhGLxZBMJiMPynVdJJPJtjmxgmVZbbmBw15R+wcg059t9MEw22m3+8m2bWQyGcTjcVknlUo91neLRXaSs2fPBs4ZMX+7Wq3KTBxE26VaraJYLCKVSiGZTLY9IGk/WVxclHP3O7EsS14v4/E4crlcx3UC/dYnIupkenoaExMTOHPmTCDtrYgPRLwpYrpisRi45mQyGZw5cwaTk5NtiRH6ps6F6YfjOJ6maXL+Yz6fD31Uudd65HnUNiHqEe2O48g5l+qjswXxqPWw9++23e4n8SjzbDYr97u6uuppmvbYP05czGNW5yOn0+nIc41oq/jXSPj//9xvHMfx8vm8bGsn4vomrpfi+mUYhlrV8zZQn4ioExGjqWuIvNY6K03TZHxQr9dl3CXWJ/k5jiPXEW1U5ytmF7Ozs3JRmNc6IP8FUxBBtFquEhdYNQD1fPuOCp7y+bxcYDVoF+jd7qd0Oh34fKFUKj32gToRBWWz2a7XoFKp1PMvntXVVU/X9cAX4ijiGqjuW/wiVI+r3/pERJ2I+MqfuMAv7Loi4q6oa1u3fXazqakvYjqHIPIXP3jwIFAvl8u1pUTrVyKR6PgUxZmZGfzhD3+AaZqoVquhqc92y2730/z8fOjDdzKZDJaWltRiInrMHTt2LHJanG3bOHbsmFoc6eLFi/j4449hWVbXZwt89tlnAIBf//rXgXKRQ/+TTz4JlPdbn4iok4sXL0LX9chnqXieFxmj6bquFgGtNW+GYeCtt95SN/VkU4F6WDCcSCQC/15cXES5XMb169cD5f0YGRlpm3PtJ+Y9jo+P47e//S3gu4D3SiwaUOcc+VmWJed453I5mRc5al64sNv9ZBgGqtVq6DYiIr/r16/DNE0cO3as7Zph2zaOHj2KdDrd87VqaWmp54d13blzBwDws5/9LFAunja7vLwcKO+3PhFRFNu2US6XcfLkSXVTR+JZGZ0SAfzxj39EvV7f0BrKTQXqYY/hVh8S8tZbb6FQKEQ+rbAb27blA0Ci3Lx5E+l0GgDw+uuvAwBWVlaUWtEWFxdx5MgRxONxOI4Dx3EwNjaGqampwCKATCaDW7duAQDW19eRy+Xw5ptvRj4SXNjtfrpw4QIAYGJiou3LRy8qlUrbYtVur418DhENhg8++ACGYWBiYkL+YhFBuq7rePfdd9W3bAlxXVT/cipubDSbzUB5v/WJiKJ8+eWXAIDDhw+rm0ItLi4il8vh6tWrWFhYiLzTDgC//OUvAQDlclnd1J06F6Yf4uEsfv55OPl83tN13XMcx6vVal0f/iLm8YS9ouami7lB/kn/4gEmvS7M0jStrR2ebz/+fYtjFO3yfAvCoo5xEPqpVCrJ+aH+ha1ERGEcx/EMw5DXC03TPMMwer6uhhHXtijiOhYmbFtYmdBpGxGRSqxz7HaNU2OwdDrd0/xztB7c169NXcXUxY/+gFQEr+Lgdd9TAUutJ/n5n6TnRSyS7BYEz87Oti0eFdkFeglGxWeqi5E8XzYB/5MfRf1+OnsQ+snzZV4QAbtpml1PyO2CkC8ZfPHF1868eiWCdSg3JzaKgToRDapu16cw/qd+d4s5xXW0X5ua+jI0NIRSqYRz584hFovh9u3b+PjjjwEAU1NTME0T4+PjWFxcRL1el7koM5kMdF2X83o6GR4ehmEYarF09epVVKvVwLSLy5cvAwD+/e9/q9XbfPHFFwCA5557Tt0k/fjjj2pRXwahn9D6c/ClS5dQrVZhGAbK5TLOnz+vVtsRrS+JfPHF1y68NmJ9fR0PHz5Ui7eUWPTeaw70fusTEW2l8fFxzM/PA61F+N2uRfV6XS3qalOBOlrB5NraGjzPw927dzE8PAzLsgILI7/55pu2IDKZTOLevXuBsih3795tm4OI1pzJer2Oer3e9otI07TIrAW7YTf7STU0NISlpSXouo6ZmRl1cxvOUSd6/Liui1QqhXq9jlKpBLQWRHX7RbQZIivMt99+q24CWgvj/fqtT0S01YaGhuRaxahr0WZsOlBXua6LXC7XtjBSzXICoO/Hs8fj8UAWgrm5ORiGEboAc2xsDM1mE4uLi+qmgJdeegkAcP/+fXWT9OSTT6pFm7aT/RQmkUgEUkZ2Mjo62vZFqNvL/yQvItpb/EH6ysoKMpkMVlZWUK/XkUqlti1YP3jwIAC03bkX17ORkZFAeb/1iYh2U1QKx062PFD/29/+hng8jhMnTgTKwy7sYQF2lEqlgmazKdNuua4Ly7LwxhtvqFUBAEePHgUA/Oc//1E3Bbz88stAK/Wieoy3b98GfJlkttJO9RMAxGKx0BSRrutu6KQhov3NH6SL1IrDw8PbHqz/6le/AkLyn4spiiI/utBvfSKiKCJDX9S1TcwuUG+Euq6Lr7/+GpqmdZ3V0OsNUr8tDdRt28bly5dx7dq1wJ3hZ599FtVqNdD4tbW1tty64uKqcl0XZ8+eBXx3nN977z00m83I+ePiLvjMzExkp6O1v0KhgGazifPnz8u609PTqFarbXe8Hz16BHQYyF7sZD8J6XRapllzXRdzc3OoVqsd834S0eMnl8u1BenC8PAwbt26hWq12vf6Ftd1sba2BvjueKtGR0eRTqcxPz+Pubk5oFX3ypUryGazbb8E+61PRBTlF7/4BdDD9JWzZ8/Km5+NRgOnT59Gs9mU6bvDiGue+CtgX9TVpZthmmZoNhTHcTxN07x8Pu95vlSB/mwmIsNJt5fnezy0eIn9Ru1L07RAisUwpVJJrtwF4BmG0ZZup1aryYwp2ODqXW8H+0mYnZ31TNMMHLtpmm3to974z79OWXb850vYeO+mXtuw0/wr6KFkNqKdkc1mu54TpVIpNFNWmHw+L7Mp+F+GYYT+f6Fmp/JnwgrTb30iojAie54aUwqO44TGU+l0uus1c3Z21oOS7rtXWxaoR6USFBYWFmRecl3XuzZqv2I/7Q8i0O02PiI1Z1hAslkbSSXl12sbdor4Ylqr1TzHcbY94ApLc0pERI8v0zQ3fBO2E8MwNrzfLZv6kslk4Hle5Hzq8fFxmfVkbW3tsf2TJPtpY3K5HGKxmFrct0ajsSVZaTql8/SLGudB0GsbhK0agyi5XA6HDh3C8PAwEokE1tbWBn5RcjKZRCqVUouJiGgPOnfuHOr1+pZmDaxUKqhWq7h27Zq6qSdbFqgTbaewxbAb8f3336tFe9bS0hK8DebE3oitGoMwjUYDzWYTr732mrpp24hsRpv5MrCRnLhERDSYRkdHkc/nkcvl5Lq+zXBdFxMTE8hmsxgfH1c394SBOg28RqOBcrmsFm/Ihx9+qBZRD7ZyDMLsxS9QW3nHhYiIBsOlS5eQzWZx9OjRrim+O6lUKnj++eeRyWTk83I2goE6DTTLsgIPLfE/VMmfOaJSqSCTychtyWSybXrLyMiIfMDT1NSUrOufulCpVJDL5ZBMJhGLxRCPxzE9Pe3by8Y1Gg05fSQWi0XuV9SLx+OyLSKjhTAyMhLaDwh5v/pS+wWtfhZtTiaTgTsJvY5BGMuyAsc6MjLSduGzLAuTk5OAMi5hLMuS7crlcmg0GnLc4/G4bFulUkEqlZLlalDt349//P195k+j5S+3bRvT09M4duwYAKBcLgf6BK0pMWHtCCu3bTvQ967rolgsys/0j5d6no+MjHQdAyIi6s+lS5fw6aef4ubNm6G/M7uxLAv/+Mc/8Omnn24qSAc2sxKNaAeJ1dVhFhYWPABeNpv1HMfxHMeRCyXVzBSFQiFyAaF4j9hWr9flwl41Q47YTy8LMcXCYNM0vXq97jmO46XTaQ+ANzs7G6hbr9c9TdNkWzzfcanHILKj+I9BLMI0DEMuWBarzdUFraINhmHI4xALLDVNC9T1uoxBmHw+H2hjvV6Xx1wqlQJ1+1nYKcbbNE0vn8/LMRf7zmazsv/8Y6gu4PbvRxD9r/brwsJCW/Yoccxqv3qtcQjrr27lIhPKwsKC7D/RJ+J4RX+KNquZoYiIaP/o/bcu0S4KC24ETdNCV1OLAM0fXHUK1MNWe4sgV63fb6CuaZoMvD1fkJdOpwN10+l0W13PF+z7iawv/mMQWYXUQFgEsX5RfSH2q6aR6jQGKpHmSv2iJLLgqF8E+gnURV21P0Tb1cBZtFP9ohMVaItjF+Xiy4861lHvF6L6q1u5v981TZN9omla22eJY1D7mYiI9gdOfaE9rdJ6EuvY2Ji6CSdPngQAfPnll+qmUEtLS/KBLMKLL74Y+PdGHTp0KPAQKpHN54cffvDVAubn59vqAoBpmj0tXHzw4AEA4JlnngmUq/vzO3z4cODfYkHnTz/9FCjvh5jP/uqrrwbKh4aGYJomms3mphfqqE94E22OWpD6zTffqEWhhoeHMTs7i3K5jGKxiOPHj+PixYs7loHJ/5Ch9fV1vP322/I8V9smjmk7F/oSEdHuYaBOe5p4SmunVINRT68dNGKusTrnORaLycC323zkZ599FgDw8OHDQLnrutA0LVC2nT7//HMg5AuD32a+CGy3U6dOIZ1OY2pqCgcOHEAmk1Gr7Chxnvvn8Pvnum/nQl8iIto9DNSJfCqVCqanp5FKpRCPx3HkyBG1yrYrFApoTUtre3W7q/v6669D0zScO3dO3mWdm5tDtVrFO++8o1anDnRdBwAsLy/DdV11865YXV1tOyfEi4iI9h8G6rQnRN0NfumllwAA9+/fVzdJTz75ZOjPqlwuh4mJCfz85z/HBx98gPX1dayurqrVts0LL7wAALh37566qWeJRAK3bt3C+vo6dF1HLBbD1atXUSgUNpUvHB3GIMzBgweBkDv7fk888YRaNDAsy8Ly8jJqtRqazSaOHz+uVul6/CLQ3wriPP/vf/+rbiIion2MgTrtCYcOHVKLAAAvv/wy0Aqs1Luet2/fBlp3mYWoOee2bWNmZgbZbBaZTEbO6e4UaG61RCIBwzA2dQe30Wjg97//PVZWVuSd1q16wmfUGIR55ZVXAADvv/9+oNx1XZTLZei6HpiLPUhs20Yul8PS0lJgvrqaTrPb8atz6NFq/0aI8/zOnTvqJiIi2scYqNOeMDQ0BLSmcYj/WpaFRCKBQqGAZrOJ8+fPy0Boenoa1WoVhUJBvhcAnn76aQDAjRs34LquzMMt7o7Oz8/LfViWhU8++QQIuWMv/t0tkBfTT9RFqmKuuRq4XbhwAc1mE6dPn5bvbTQaKBaLbYGieO+jR49k2UcffYRms+mrFS2qDWJ+uX+/6DAGYcbHx2GaJsrlsqzvuq68M60G8OKzxJerTkRdte9EO9SxEmsU1LUKYfuxbRtHjx7FO++8I7+snTp1Cpqm4fLly23t1XUd5XIZtm3DdV2Z2x2+/hI5eBuNBs6fPy/vtPs/N+pnIZFIIJ/PywWuoo74UqEeFxER7RNqGhiiQeQ4jkwbqOt6W/rBUqkkUxCilY9aTccnlEolmSs7nU7LVIilUkmmdNR1XearFvsUKfBEXnPxUo9FqNVq8nPEPsPK1RSNCwsLbW0pFAqBlI3qMYiUfiKVoH+bfz+iT9T3izao5f5Ugd3GQOU4jlcoFALHk06n29I+inSI/jpRovpO3YcYK5FeU7zEmIbtx78Pf850tU/8ue9rtZpsn2EYbf0l8uVrvtz4og/9n+HvIzVfu5/an6Zpdh0HIiLau2IeVyER7RvFYhE3btzAV199FUjJ2Gg0kE6nUa/Xsb6+HngPERERDSZOfSHaR6ampnDy5Mm2vOlDQ0MYGxvreVoMERER7T4G6kT7iK7ruHLlChYXF2WZ67qwLEsuliUiIqK9gVNfiPaRRqOBv/zlL1heXg48ydQ0TUxOTu76g3uIiIiodwzUiYiIiIgGEKe+EBERERENIAbqREREREQDiIE6EREREdEAYqBORERERDSAGKgTEREREQ0gBupERERERAOIgToRERER0QBioE5ERERENIAYqBMRERERDaD/AbX2qOhWJcvSAAAAAElFTkSuQmCC\" width=\"746\" height=\"75\"\u003e\u003c/div\u003e\n\u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003e\u003cstrong\u003eEffect of media composition and culture conditions on EPS production\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA comprehensive experimental approach was employed to ascertain the optimal carbon source for enhancing exopolysaccharide production. This involved utilizing five distinct carbon sources, each at a concentration of 20 g/L, within a customized MRS broth medium (Table 1). The experimental design comprised\u0026nbsp;a one-factor-at-a-time (OFAT) experimental framework meticulously designed to investigate the impact of medium composition and prevailing culture conditions on the production of exopolysaccharides. No statistically significant differences (p\u0026gt;0.05) in EPS yield or cell growth were observed when five carbon sources were prepared as supplements in fermentation media. However, sucrose exhibited the highest EPS production at 85 mg/L, 1.5-fold higher than MRS without saccharide supplementation. After determining sucrose as the optimal carbon source, different sucrose concentrations (25\u0026ndash;125 g/L) were examined to ascertain the appropriate sucrose concentration for EPS production. The preliminary screening revealed optimal EPS production was achieved at 150.1 mg/L with 50 g/L sucrose supplementation.\u003c/p\u003e\n\u003cp\u003e[Table 1]\u003c/p\u003e\n\u003cp\u003eA culture was also cultivated on a water bath shaker to ascertain the impact of agitation on EPS production. The agitation speed of the water bath shaker was adjusted within the range of 0 to 150 rpm and the quantity of produced EPS was examined after 48 hours. The application of an agitation speed of 100 rpm was found to enhance EPS production by up to 2-fold compared to fermentation conducted without agitation.\u0026nbsp;A statistically significant difference (p\u0026lt;0.05) was observed between cultures cultivated with and without agitation (Figure 1). Agitation speeds exceeding 100 rpm during fermentation did not lead to a significant enhancement in EPS production.\u003c/p\u003e\n\u003cp\u003eA preliminary study was conducted to evaluate the effect of inoculum size on EPS production using inoculum concentrations of 2%, 4%, 6%, 8%, and 10%. Fermentation was carried out at an agitation speed of 100 rpm, and EPS was harvested after 48 hours. The results indicated that the highest EPS yield was obtained with an inoculum size of 6%, reaching 204.9 mg/L\u0026mdash;approximately 1.2 times higher than that obtained with lower concentrations. Increasing the inoculum size beyond 6% did not result in a significant enhancement of EPS production.\u003c/p\u003e\n\u003cp\u003e[Figure 1]\u003c/p\u003e\n\u003cp\u003eFermentation time is critical in producing exopolysaccharides (EPS) due to different microorganisms varying growth phase rates. In this screening stage, the determination of the optimal fermentation time was carried out by culturing with a 6% starter concentration, 50 g/L sucrose supplementation, and an agitation speed of 100 rpm for 8, 16, 24, 32, 40, 48, and 56 hours. The results showed that the highest EPS production was achieved at 24 hours of fermentation. Extending the fermentation time did not result in a further increase in the number of EPS produced. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eOptimization and validation of EPS yield\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;Following the implementation of a screening process employing the OFAT method, sucrose (X1), inoculum size (X2), and agitation speed (X3) were identified as the variables to be investigated for their capacity to enhance the production of EPS from \u003cem\u003eP.\u0026nbsp;\u003c/em\u003e\u003cem\u003epentosaceus\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003eSL4. The optimal levels of these variables were determined by utilizing a Box-Behnken design, a statistical design methodology within the response surface methodology (RSM) framework. This experimental design involved the execution of three replicates, encompassing twelve factorial points and five central points (Table 2). The measurement of the resultant EPS yield served as the response variable in this study. Therefore, a quadratic model was developed for EPS yield as represented below in terms of core values:\u003c/p\u003e\n\u003cp\u003e\u003cimg 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β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\" width=\"746\" height=\"89\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eX\u003csub\u003e1\u003c/sub\u003e is sucrose concentration, X\u003csub\u003e2\u003c/sub\u003e is agitation speed, and X\u003csub\u003e3\u003c/sub\u003e is inoculum size.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e[Table 2]\u003c/p\u003e\n\u003cp\u003e[Table 3]\u003c/p\u003e\n\u003cp\u003eThe triplicate responses from each run were analysed using one-way ANOVA (Table 3). The quadratic polynomial model derived from Response Surface Methodology (RSM) effectively characterized the relationship between sucrose concentration (X₁), agitation speed (X₂), and inoculum size (X₃) on exopolysaccharide (EPS) production (Figure 2). ANOVA results confirmed the statistical significance of the model (F-value = 14.56, p = 0.001), validating the inclusion of linear, interaction, and quadratic terms. Regression coefficients revealed significant negative linear effects of X₁, X₂, and X₃, as well as notable antagonistic interactions between the factors (X₁X₂, X₁X₃, and X₂X₃), suggesting synergistic modulation of EPS synthesis. The quadratic terms were also significant, with inoculum size (X₃\u0026sup2;) exhibiting the greatest curvature effect, thereby delineating an optimal response surface. Optimization analyses, supported by three-dimensional response surface plots, identified the optimal fermentation conditions as approximately 45\u0026ndash;50 g/L sucrose, 90\u0026ndash;100 rpm agitation, and 6% inoculum size, corresponding to maximal EPS yield. These findings highlight the critical influence of both individual and combined fermentation parameters and emphasize the necessity of multifactorial optimization in maximizing EPS biosynthesis efficiency.\u003c/p\u003e\n\u003cp\u003eThe optimal value, predicted using the BBD experimental model, was selected for the verification test. The verification test involved four repeated experiments by setting sucrose, agitation speed, and inoculum size at predicted levels: 50 g/L, 100 rpm, and 6%, respectively. The results showed no significant difference (p\u0026gt;0.05) between the experimental and predicted values, suggesting that the model can predict the EPS yield.\u003c/p\u003e\n\u003cp\u003e[Figure 2]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe molecular weight of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe molecular weight of EPS was estimated by the polyethylene glycol (PEG) as standard, which has a known molecular weight. Gel permeation chromatography was used to fractionate the crude EPS into two components eluted by deionized water. The polysaccharide content was detected at 94.81% (EPS-1) and 5.19% (EPS-2), indicating that the production of EPS from\u0026nbsp;\u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 after purification was approximately 405.79 mg/L (peak 1) and 22.21 mg/L (peak 2). The presence of sharp, single, and symmetrical peaks in gel permeation chromatography suggests that EPS is a homogeneous fraction. The molecular weight of the exopolysaccharide (EPS) was estimated to be 7.35\u0026times;10\u003csup\u003e5\u003c/sup\u003e KDa for EPS-1 and 2,7 KDa for EPS-2.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunctional group analysis of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFT-IR spectroscopy has been used to investigate the presence of functional groups qualitatively and the linkage among the monosaccharides of the EPS. The absorbance was recorded using FT-IR at a wavenumber from 4000 to 500 cm\u003csup\u003e-1\u003c/sup\u003e. The IR spectrum of the EPS from\u0026nbsp;\u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 revealed absorption characteristics for polysaccharides, which had many peaks from 3406 to 581.19 cm\u003csup\u003e-1\u003c/sup\u003e. A broad peak around 3406 cm\u003csup\u003e-1\u003c/sup\u003e corresponds to O\u0026ndash;H stretching vibrations, which are typical of polysaccharides due to the abundance of hydroxyl groups in sugar monomers. The peaks at approximately 2920 and 2850 cm⁻\u0026sup1; are attributed to C\u0026ndash;H stretching vibrations of aliphatic \u0026ndash;CH₂ and \u0026ndash;CH₃ groups, indicating the presence of alkyl chains. A distinct peak at 1653 cm\u003csup\u003e-1\u003c/sup\u003e is attributed to the C=O stretch bond vibration. In addition, the weak absorption peak observed at 1542 cm\u003csup\u003e-1\u003c/sup\u003e is related to the N-H. A weak absorption at 1417 cm\u003csup\u003e-1\u003c/sup\u003e is related to the presence of C-H, while intense absorption peaks at around 1219 and 1053 cm\u003csup\u003e-1\u003c/sup\u003e are assigned to carboxylic acid and ester groups C-O stretching vibrations. \u0026nbsp;Several minor peaks associated with \u0026alpha;- or \u0026beta;-glycosidic configurations and ring vibrations of polysaccharides were also observed in the absorbance range of 914\u0026ndash;581 cm⁻\u0026sup1;. Overall, the spectral features confirm the presence of hydroxyl, carbonyl, and glycosidic functional groups, supporting the identification of the sample as a complex exopolysaccharide.\u003c/p\u003e\n\u003cp\u003e[Figure 3]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMicroscopic manifestation of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eScanning electron microscopy (SEM) is a technique for illustrating common physical properties, including microstructure, surface morphology, and texture of EPS. As illustrated in Figure 4, the SEM of EPS produced by \u003cem\u003eP.\u0026nbsp;\u003c/em\u003e\u003cem\u003epentosaceus\u003c/em\u003e under optimal conditions (sucrose concentration of 50 g/L, agitation speed of 100 rpm, and inoculum size of 6%) with incubation at 37\u0026deg;C for 24 h displays a compact and porous structure with a flake-like morphology, as well as irregular, overlapping aggregates of varying size.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe Energy Dispersive X-ray (EDX) has become a staple quantitative elemental sample analysis method. The EDX analysis revealed that carbon and oxygen were the predominant elements, with a mass ratio of 48.33% and 42.72%, respectively, suggesting EPS is comprised mainly of carbohydrates. Heteroatoms such as phosphorus (P) and nitrogen (N) were detected at approximately 0.29% and 7.91%, respectively. In addition, the analysis revealed traces of sulphur (0.21%) in the EPS.\u003c/p\u003e\n\u003cp\u003e[Figure 4]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThermal properties of EPS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe thermal properties of the EPS were found to be critically important for the intended application.\u003cbr\u003e\u0026nbsp;Differential Scanning Calorimetry (DSC) was employed to analyse the thermal behaviour of the sample, including melting temperature, glass transition temperature, and crystallization characteristics.\u003cbr\u003eThe thermal characterization of the EPS revealed the presence of both endothermic and exothermic transitions, observed within the temperature range of 30\u0026ndash;400 \u0026deg;C for the EPS derived from \u003cem\u003ePediococcus pentosaceus\u0026nbsp;\u003c/em\u003eSL-4. The DSC thermogram of the EPS exhibited a broad endothermic peak, beginning at 46.8 \u0026deg;C and ending at 127.8 \u0026deg;C, with a melting point peak observed at 91.7 \u0026deg;C. The enthalpy of fusion (\u0026Delta;H) was calculated to be 232.2 J/g. A second endothermic peak was detected at 226.1 \u0026deg;C, with an associated enthalpy change of 29.21 J/g, corresponding to partial melting or the onset of thermal degradation. No detectable glass transition temperature was observed in the DSC thermogram of the EPS sample. Finally, a mild exothermic drift beyond 250 \u0026deg;C was observed, suggesting the initiation of degradation of the polymer backbone.Top of Form\u003c/p\u003e\n\u003cp\u003e[Figure 5]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTechnological Properties\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMicrobial polysaccharides have been identified as a promising renewable source for hydrocolloids in the food industry, owing to their distinctive functional characteristics, including high solubility and substantial water-retention capabilities, and act as stabilizer. In this study, the findings indicate that the water solubility index and water holding capacity of EPS were 21.53 \u0026plusmn; 0.58% and 130.52 \u0026plusmn; 1.44%, respectively. Thereby to preventing disintegration into their native phase and thus enhancing the stability of food products, in this study the probable effect of EPS concentration on suspension stability against virgin red palm oil was investigated. Emulsification activity (EA) values were tested by adjusting EPS at several concentrations (0.2-1.0%). A statistically significant difference (p\u0026lt;0.05) in the EA values was observed when the emulsion was prepared with concentrations ranging from 0.2 to 1% (Figure 6). Therefore, EPS at 0.6% is the optimal concentration for EPS when used as a stabilizer. No significant difference in its emulsifying capacity is observed at higher concentrations. The stability of the emulsion was determined by allowing it to sit for a more extended period, up to 72 hours, after which the emulsion gradually decreased after 24 hours. A common challenge in applying emulsifiers is maintaining the emulsion formed under extreme conditions, such as high and low temperatures and varying pH levels. This study evaluated the emulsion-forming and stabilizing properties at varying temperatures (0, 30, 60, and 90\u0026deg;C) and pH levels (3-7). The emulsion of EPS with virgin red palm oil exhibited stability within the pH range of 4-7 and emulsification stabilizer declined at storage temperature above 30\u0026deg;C.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e[Figure 6]\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eSourdough prepared using tomi-tomi fruit (\u003cem\u003eFlacourtia inermis\u003c/em\u003e) starter is a source of various lactic acid bacteria species (Kemhay et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). \u003cem\u003ePediococcus pentosaceus\u003c/em\u003e SL4, one of the lactic acid bacteria successfully isolated, has the potential to produce a considerable number of exopolysaccharides. In the screening process using MRS Agar medium supplemented with 0.1% Congo Red indicator, \u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 demonstrated the ability to produce exopolysaccharides, as indicated by a color change of the MRS Agar medium to dark brown. Several studies have also reported similar findings, where \u003cem\u003eP. pentosaceus\u003c/em\u003e was successfully isolated from sourdough made with grape, apple, and pomegranate starters (Gord\u0026uacute;n et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Ripari et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The exopolysaccharides produced by certain lactic acid bacteria during dough fermentation have also been reported to enhance the sensory characteristics of sourdough (Zhang et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023b\u003c/span\u003e). Other research suggests that exopolysaccharides hold potential as functional food ingredients or additives to improve the physical and chemical properties of food products (Loeffler et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Those potentials can be further explored by enhancing \u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 to produce EPS then characterize them.\u003c/p\u003e\u003cp\u003eMany factors contribute to the enhanced production of exopolysaccharides (EPS) by lactic acid bacteria. The presence of an excess carbon source in the form of saccharides in the fermentation medium can enhance the ability of lactic acid bacteria to produce exopolysaccharides. This phenomenon is associated with an increase in osmotic pressure due to the high sugar content. Several studies have shown that the maximum EPS yield (53.79 g/L) was attained with \u003cem\u003eLeuconostoc mesenteroides\u003c/em\u003e DRP105, utilizing sucrose at 86.83 g/L as a carbon source (Xing et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). A parallel finding was reported for \u003cem\u003eLeuconostoc citreum\u003c/em\u003e B-2, which yielded 59.33 g/L with sucrose supplementation at 99.8 g/L (Wang et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). \u003cem\u003eP. acidilactici\u003c/em\u003e MT41-11 was reported to be able to produce exopolysaccharides of 500 mg/L when fermented MRS media by adding 5% sucrose (Bai et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). However, other studies have acknowledged that alternative carbon sources, such as maltose and lactose, can stimulate EPS production by \u003cem\u003eEnterococcus\u003c/em\u003e and \u003cem\u003eLactobacillus\u003c/em\u003e (Imran et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The variation in EPS production can be attributed to differences in bacterial strains and the type of carbon source utilized (Imran et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Each strain has a different metabolic pathway, so it uses different carbon sources. High sucrose concentrations in media induced the β-fructofuranosidase enzyme production from microorganisms to catalyse the conversion of sucrose into glucose and fructose, which can later be used as substrates (Sheng et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn addition to the critical role of carbon source selection and concentration, agitation as a parameter of the fermentation process has been identified as a factor that also significantly affects EPS production. Agitation also plays a role in maintaining the homogeneity of the medium throughout the fermentation process. The inadequate mixing of high-viscosity media gives rise to non-homogeneity. Nevertheless, elevated mixing speed engenders elevated shear stress in the culture medium, diminishing the EPS yield (Prasad and Purohit \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Fermentation of \u003cem\u003eL. rhamnosus\u003c/em\u003e in a mixed medium of fructose, glucose, and saccharose at 20 g/L with an agitation speed of 80 rpm, which exhibited augmented EPS production in comparison to fermentation conducted without agitation (Oleksy-Sobczak et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Furthermore, previous studies have also investigated the optimization of exopolysaccharide production through the regulation of inoculum concentration; however, inoculum concentrations exceeding 10% did not result in a significant improvement in EPS yield from an economic standpoint (Oleksy-Sobczak et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBeyond agitation and inoculum concentration, fermentation time emerges as a critical parameter affecting EPS yield. Precise determination of the optimal fermentation duration for each microorganism is vital, as mismatches between the growth and EPS production phases can compromise both efficiency and yield. Extended fermentation periods often result in decreased EPS production, largely due to enzymatic degradation by glycosyl hydrolases (Haj-Mustafa et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Amiri et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), including rhamnosidase produced by species such as \u003cem\u003eLactobacillus plantarum\u003c/em\u003e (Ferreira-Lazarte et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and \u003cem\u003eLactobacillus acidophilus\u003c/em\u003e (Mueller et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This pattern aligns with observations from Haj-Mustafa et al. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), who noted peak EPS synthesis by \u003cem\u003eL. rhamnosus\u003c/em\u003e 519 and \u003cem\u003eL. acidophilus\u003c/em\u003e LA5 within the first 49 and 24 hours, respectively, followed by gradual declines. Similarly, \u003cem\u003eEnterococcus mundtii\u003c/em\u003e A2 demonstrated maximum EPS yield at 16 hours, with reduced production at longer incubation times (Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In the RSM optimization, by adjusting sucrose concentration, agitation speed, and inoculum concentration to optimal conditions, the EPS yield reached 428.0 mg/L, which is 7.8-fold higher than the original yield (55 mg/L in MRS medium, Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Moreover, this yield was also higher than those reported for \u003cem\u003eEnterococcus mundtii\u003c/em\u003e A2 (382.27 mg/L) (Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), \u003cem\u003eLactobacillus plantarum\u003c/em\u003e NTMI05 (197 mg/L), and \u003cem\u003eLactobacillus plantarum\u003c/em\u003e NTMI20 (187 mg/L) (Imran et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). However, some other species such as \u003cem\u003eLeuconostoc lactis\u003c/em\u003e L2 (58.1 g/L) (Jiang et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), \u003cem\u003eLactobacillus plantarum\u003c/em\u003e KX041 (599.52 mg/L) (Wang et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), and \u003cem\u003eEnterococcus faecium\u003c/em\u003e MC-5 (16.48 g/L) (Tilwani et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) have been reported to produce even higher amounts of EPS. In addition to strain differences, variations in fermentation optimization parameters can also influence the EPS yield obtained.\u003c/p\u003e\u003cp\u003eThe molecular weight of the first fraction of EPS is higher than that of other EPS produced by coccus genera of lactic acid bacteria, which range from 2.5\u0026times;10\u003csup\u003e1\u003c/sup\u003e KDa to 2\u0026times;10\u003csup\u003e3\u003c/sup\u003e KDa (Jia et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Bai et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Jiang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, it is nearly equivalent to the molecular weight of dextran, which is produced by fructophilic lactic acid bacteria, \u003cem\u003eApilactobacillus waqarii\u003c/em\u003e strain HBW1 (Ahmad et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These discrepancies in molecular weight may be attributable to inter-species and inter-strain variations. The elevated molecular weight of EPS produced by lactic acid bacteria renders them a viable alternative to stabilizers in the food industry, biomedicine, and pharmaceuticals.\u003c/p\u003e\u003cp\u003eThe FTIR spectrum of the EPS in this study aligns with previous findings. A broad band at 3590\u0026ndash;3389 cm⁻\u0026sup1; corresponds to O\u0026ndash;H stretching, indicating abundant hydroxyl groups typical of polysaccharides (Amiri et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). This region is also associated with water solubility (Vinothini et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The peak at around 2932 cm⁻\u0026sup1;, attributed to C\u0026ndash;H and C\u0026ndash;O stretching, matches that reported in EPS from \u003cem\u003eL. plantarum\u003c/em\u003e KX041 (Wang et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). A peak at 1633 cm⁻\u0026sup1; corresponds to C\u0026thinsp;=\u0026thinsp;O stretching, as also found in EPS from \u003cem\u003eL. acidophilus\u003c/em\u003e LA5 (Imran et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). A weak absorption peak at 1542 cm⁻\u0026sup1; is related to N\u0026ndash;H bending of amide II from proteins (Wang et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Notably, absorption in this region was not observed in EPS from other \u003cem\u003eP. pentosaceus\u003c/em\u003e strains (Jiang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The fingerprint region (800\u0026ndash;1200 cm⁻\u0026sup1;) shows typical polysaccharide features, with 1200\u0026ndash;1000 cm⁻\u0026sup1; indicating C\u0026ndash;O and C\u0026ndash;O\u0026ndash;C glycosidic linkages (Amiri et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). A band at 813.44 cm⁻\u0026sup1; suggests α-glycosidic linkages (Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). These features confirm that the EPS contains characteristic functional groups commonly found in microbial exopolysaccharides.\u003c/p\u003e\u003cp\u003eMicroscopic analysis revealed that the morphology of exopolysaccharides (EPS) produced by different bacterial strains can vary significantly, even among those belonging to the cocci group. For example, levan produced by \u003cem\u003eEnterococcus faecium\u003c/em\u003e MC-5 exhibits a uniform spherical morphology (Tilwani et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), whereas glucan synthesized by \u003cem\u003eEnterococcus hirae\u003c/em\u003e KX577639 presents an irregular and interconnected structure (Jayamanohar et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Such variations in EPS microstructure are primarily attributed to differences in the composition of monosaccharide units, chain length, and the types of glycosidic linkages forming the polymer backbone. Furthermore, EPS characterized by porous and spongy textures, as observed in this study, possess enhanced water-binding capabilities, which may contribute to their functional properties. This property that lends itself to applications in the food industry as a gelling, thickening, and stabilizing agent. This characteristic renders it a promising candidate for producing edible films and coatings for food products (Jiang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The heteroatom P and N in the partially purified EPS \u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 proposed a trace of phospholipids and protein, respectively (Zaghloul and Ibrahim \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Also, the analysis revealed traces of another element, including sulphur, which were close to the EPS produced by \u003cem\u003eLeu. mesenteroides\u003c/em\u003e DRP105 (Xing et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2018\u003c/span\u003e)d \u003cem\u003esakei\u003c/em\u003e L3 (Wang et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The elemental composition of the SL4-EPS was similar to the EPS produced by \u003cem\u003eEnterococcus\u003c/em\u003e sp.BE11 (Zaghloul et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) and \u003cem\u003eEnterococcus mundtii\u003c/em\u003e A2 (Zanzan et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe DSC analysis of the exopolysaccharide (EPS) sample demonstrated thermal behaviour consistent with previous studies on similar biopolymers. This analysed was employed to ascertain the correlation between physicochemical properties and temperature during application (Vinothini et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The prominent endothermic peak, attributed to the loss of bound water, which is facilitated by the abundant carboxyl groups in EPS that bind with water molecules. As the temperature rises, the hydrogen bonds that bind the water molecules weaken, releasing them (Tilwani et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). This is in line with the study that has been conducted by Jiang et al. (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), who observed dehydration peaks in EPS from \u003cem\u003eP. pentosaceus E-8\u003c/em\u003e occurring at 82.6\u0026deg;C due to the hygroscopic nature of polysaccharides. The relatively high enthalpy value in first endothermic peak in this study suggests a significant water-binding capacity, which is critical for the material\u0026rsquo;s physical properties and stability. The second endothermic phase occurred between 226 and 233.8\u0026deg;C, corresponding to the higher protein content in the EPS. Higher temperatures associated with higher energy requirements are necessary to remove proteins covalently bound to polysaccharides (Wang et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Studies on microbial EPS have demonstrated thermal stability up to approximately 220\u0026ndash;260\u0026deg;C before degradation initiates, consistent with the mild exothermic transition observed here beyond 250\u0026deg;C. For instance, EPSF2 from \u003cem\u003eEnterococcus\u003c/em\u003e sp. F2 maintained structural integrity up to 266.6\u0026deg;C (Jiang et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and EPSE8 from \u003cem\u003eP. pentosaceus\u003c/em\u003e exhibited stability up to 257\u0026deg;C in TGA analysis (Jiang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The slight variation in peak temperature and enthalpy values may be attributed to differences in molecular weight distribution, degree of branching, and molecule structure (Lakra et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Jiang et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSolubility, a pivotal metric in assessing the saturation concentration of EPS in an aqueous environment at a designated temperature, is a critical factor in determining the textural characteristics of food products (Jiang et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Water-holding capacity is the ability to hold an immense amount of water (Vinothini et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The water solubility of EPS in this study was nearly higher than that of EPS extracted from \u003cem\u003eLeu. lactic\u003c/em\u003e KC117496 (14.2%), yet the water-holding capacity value obtained was lower (Saravanan and Shetty \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In contrast, D-glucan produced by \u003cem\u003eEnterococcus hirae\u003c/em\u003e KX577639 exhibited a water solubility index of 46.5% and a water holding capacity of 202.04%, which is higher than the results obtained in this study (Jayamanohar et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The molecular weight and percentage of branching appear to influence their water solubility (Jayamanohar et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Furthermore, the high water-holding capacity of EPS can be attributed to its porous polymer structure, which enables water retention through hydrogen bonds (Saravanan and Shetty \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The elevated WHC of EPS indicates its capacity to retain substantial amounts of water, thereby mitigating the dehydration levels of the product. A previous study has demonstrated that incorporating 4% EPS into minced meat can stabilize the meat during cooking, thereby reducing losses (Wu et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). EPS's high solubility and water-retention capacity render it a suitable stabilizer and bio-thickener. Xanthan gum polymer has been reported to have a solubility index of 64.86% and a water holding capacity of 1000.33% (Sarkar et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), respectively, and has been recommended by the US as a stabilizer, thickener, binder, and texturizer. EPS from \u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 has been identified as a potential alternative renewable hydrocolloid despite xanthan gum's higher solubility and water binding.\u003c/p\u003e\u003cp\u003eThis finding is consistent with the observations reported for EPS derived from \u003cem\u003eL. plantarum\u003c/em\u003e ATCC 8014, which exhibited emulsification activity of approximately 60% at 72 hours (Balyan \u003cem\u003eet al.\u003c/em\u003e, 2024). However, a separate study demonstrated that the emulsifying properties of a mixture of lactic acid bacteria found in yogurt could stabilize limonene emulsions by over 70% after 72 hours (Balyan et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024b\u003c/span\u003e). Notably, the optimal emulsifying capacity of EPS produced by \u003cem\u003eP. pentosaceus\u003c/em\u003e SL4 is about 74.75% when employed in products with a temperature of 4\u0026deg;C and a pH range of 4\u0026ndash;7. Furthermore, a comprehensive study established that a reliable emulsifier must retain at least 50% of the original emulsion volume after 24 hours of its formation (Vinothini et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eConflict of Interest\u003c/h2\u003e\u003cp\u003eThe authors declare no conflicts of interest related to the results of this study with other parties\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eDAP: study design, data acquisition, data analysis and interpretation, drafting the manuscript; RY: material support, critical revision of the manuscript; CH: study design, study supervision, critical revision of the manuscript; DAS: study concept and design, study supervision, data interpretation, critical revision of the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThis study was supported by the Indonesia Endowment Fund for Education Agency (LPDP) and the National Research and Innovation Agency (BRIN).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAhmad W, Boyajian JL, Abosalha A, et al (2022) High-Molecular-Weight Dextran-Type Exopolysaccharide Produced by the Novel Apilactobacillus waqarii Improves Metabolic Syndrome: In Vitro and In Vivo Analyses. 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Int J Biol Macromol 192:716\u0026ndash;727. https://doi.org/10.1016/j.ijbiomac.2021.10.047\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"491\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"4\" valign=\"bottom\" style=\"width: 491px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1\u0026nbsp;\u003c/strong\u003eEffect of carbon source on cell growth and EPS produced by \u003cem\u003eP. pentosaceus\u0026nbsp;\u003c/em\u003eSL4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eCarbon Source\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003eSaccharide Cons (g/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003eGrowth (OD 600 nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 198px;\"\u003e\n \u003cp\u003eEPS yield (mg L\u003csup\u003e-1\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eMRS+Glucose\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.138 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 198px;\"\u003e\n \u003cp\u003e75.0\u0026plusmn;0.00\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eMRS+Sucrose\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.154\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 198px;\"\u003e\n \u003cp\u003e85.0\u0026plusmn;10.61\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eMRS+ Lactose\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.141\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 198px;\"\u003e\n \u003cp\u003e60.0\u0026plusmn;7.07\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eMRS+ Fructose\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.133\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 198px;\"\u003e\n \u003cp\u003e60.0\u0026plusmn;0.00\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eMRS+ Galactose\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.149\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 198px;\"\u003e\n \u003cp\u003e76.0\u0026plusmn;17.67\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 113px;\"\u003e\n \u003cp\u003eMRS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 76px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 104px;\"\u003e\n \u003cp\u003e0.139\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 198px;\"\u003e\n \u003cp\u003e55.0\u0026plusmn;5.67\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\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\u003e\u003csup\u003ea\u003c/sup\u003e Values in the same column not followed by a common superscript letter are significantly different (p \u0026lt; 0.05)\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"424\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\" valign=\"bottom\" style=\"width: 424px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e BBD matrix along with actual and predicted EPS yield (n=3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" rowspan=\"2\" style=\"width: 71px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"3\" rowspan=\"2\" style=\"width: 213px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eVariables\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 140px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResponse\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 140px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eEPS (mg/L)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRun\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSucrose (g/L)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAgitation (rpm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eInoculum (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eActual\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePredicted\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e344,9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e332,6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e346,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e351,2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e347,5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e359,8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e315,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e306,5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e388,8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e392,6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e305.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e318,9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e463.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e440,2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e359\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e357,7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e412,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e398,7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e396,5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e418,1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e350,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e359,1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e295,7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e290,7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e375,7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e377,0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e404,4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e400,6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e426,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e418,1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e428\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e418,1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 64px;\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 81px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64px;\"\u003e\n \u003cp\u003e419,8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e418,1\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\u003e\u003cstrong\u003eTable 3\u003c/strong\u003e. Analysis of variance for quadratic model of exopolysaccharide production\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"589\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSource\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSum of squares\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e\u003cstrong\u003edf\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean square\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 72px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 71px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eModel\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e28832.90\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e9\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e3203.66\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 72px;\"\u003e\n \u003cp\u003e14.56\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 71px;\"\u003e\n \u003cp\u003e0.0010\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 90px;\"\u003e\n \u003cp\u003esignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA-Sucrose_Conc\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2574.03\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2574.03\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e11.70\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0111\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eB-Agitation\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3858.81\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3858.81\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e17.53\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0041\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eC-Inoculum_Conc\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e554.44\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e554.44\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.52\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.1565\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAB\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1428.84\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1428.84\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6.49\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0382\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1193.70\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1193.70\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5.42\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0527\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eBC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e637.56\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e637.56\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.90\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.1325\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA\u0026sup2;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e602.28\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e602.28\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.74\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.1420\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eB\u0026sup2;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3410.41\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3410.41\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e15.50\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0056\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eC\u0026sup2;\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e13244.33\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e13244.33\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e60.18\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eResidual\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1540.49\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e220.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eLack of Fit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e903.42\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 47px;\"\u003e\n \u003cp\u003e3\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 83px;\"\u003e\n \u003cp\u003e301.14\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 72px;\"\u003e\n \u003cp\u003e1.89\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 71px;\"\u003e\n \u003cp\u003e0.2723\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 90px;\"\u003e\n \u003cp\u003enot significant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePure Error\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e637.07\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e159.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eCor Total\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e30373.39\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"566\" class=\"fr-table-selection-hover\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 144px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFit Statistics\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 422px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 144px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eStd. Dev.\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 140px;\"\u003e\n \u003cp\u003e14.83\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 145px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eR\u0026sup2;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 137px;\"\u003e\n \u003cp\u003e0.9493\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 144px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 140px;\"\u003e\n \u003cp\u003e372.71\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 145px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAdjusted R\u0026sup2;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 137px;\"\u003e\n \u003cp\u003e0.8841\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 144px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC.V. %\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 140px;\"\u003e\n \u003cp\u003e3.98\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 145px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePredicted R\u0026sup2;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 137px;\"\u003e\n \u003cp\u003e0.4913\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eAdeq Precision\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e11.2024\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\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":"[email protected]","identity":"world-journal-of-microbiology-and-biotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wibi","sideBox":"Learn more about [World Journal of Microbiology and Biotechnology](https://www.springer.com/journal/11274)","snPcode":"11274","submissionUrl":"https://submission.nature.com/new-submission/11274/3","title":"World Journal of Microbiology and Biotechnology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Box Behnken design, characterization, exopolysaccharides, food stabilizer, P. pentosaceus SL4","lastPublishedDoi":"10.21203/rs.3.rs-7555681/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7555681/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSourdough is a fermented food made by a complex and diverse consortium of microorganisms, predominantly wild yeasts and lactic acid bacteria (LAB), positioning it as a valuable source for the isolation of specific LAB strains. LAB are capable of synthesising metabolites known as exopolysaccharides (EPS) during fermentation which enhanced the quality of food product. The characteristics of exopolysaccharides which are produced by LAB vary greatly depending on structure, molecular size, and physicochemical properties. This study aims to determine the optimal conditions for EPS production and characterize the exopolysaccharide from \u003cem\u003ePediococcus pentosaceus\u003c/em\u003e SL4 isolated from \u003cem\u003eFlacourtia inermis\u003c/em\u003e sourdough. The investigation utilized a modified de Man Sharpe Rogosa medium and response surface methodology (RSM). The screened factors included monosaccharide types and concentrations, inoculum size, agitation speeds, and fermentation time. Using the Box Behnken design, the optimized conditions for EPS production were sucrose concentration of 5%, inoculum size of 6%, and agitation at 100 rpm, resulting in a maximum yield of 426.2 mg/L after 24 h incubation. The partially purified EPS had a molecular weight of 7.35\u0026times;10\u003csup\u003e5\u003c/sup\u003e kDa with a degradation temperature of 91.8⁰C. The irregular, porous, and spongy morphology of EPS was observed by scanning electron microscopy. The EPS exhibited physiological properties such as water solubility index, holding capacity, and emulsifying activity. Overall, EPS exhibits technological properties as a stabilizer in the food industry.\u003c/p\u003e","manuscriptTitle":"Optimization on production and technological properties of exopolysaccharide from Pediococcus pentosaceus SL4 isolated from sourdough Tomi-tomi (Flacourtia Inermis Roxb)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-17 11:02:39","doi":"10.21203/rs.3.rs-7555681/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-20T07:34:53+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-23T06:32:09+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-21T11:48:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"187732864840658241456693983782141605256","date":"2025-09-21T06:11:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"67750072796688634648547196097314922494","date":"2025-09-20T07:08:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"140317244777516345470718885894007447251","date":"2025-09-11T05:51:35+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-10T06:02:46+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-09T18:15:50+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-09T07:56:09+00:00","index":"","fulltext":""},{"type":"submitted","content":"World Journal of Microbiology and Biotechnology","date":"2025-09-07T10:36:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"world-journal-of-microbiology-and-biotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wibi","sideBox":"Learn more about [World Journal of Microbiology and Biotechnology](https://www.springer.com/journal/11274)","snPcode":"11274","submissionUrl":"https://submission.nature.com/new-submission/11274/3","title":"World Journal of Microbiology and Biotechnology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"bee51128-157e-4f0b-bb38-2af18ac32d2d","owner":[],"postedDate":"September 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-30T16:21:44+00:00","versionOfRecord":{"articleIdentity":"rs-7555681","link":"https://doi.org/10.1007/s11274-026-04904-2","journal":{"identity":"world-journal-of-microbiology-and-biotechnology","isVorOnly":false,"title":"World Journal of Microbiology and Biotechnology"},"publishedOn":"2026-03-25 16:10:56","publishedOnDateReadable":"March 25th, 2026"},"versionCreatedAt":"2025-09-17 11:02:39","video":"","vorDoi":"10.1007/s11274-026-04904-2","vorDoiUrl":"https://doi.org/10.1007/s11274-026-04904-2","workflowStages":[]},"version":"v1","identity":"rs-7555681","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7555681","identity":"rs-7555681","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}