Influence of environmental conditions on Nostoc sphaericum: physicochemical and functional characterization of exopolysaccharides using response surface methodology

Exopolysaccharide from psychrotrophic Arctic glacier soil bacteriumFlavobacteriumsp. ASB 3-3.

Microbial exopolysaccharides (EPSs) are valuable extracellular macromolecules secreted as capsules or slime layers. Various microorganisms, including bacteria, yeasts, fungi, and algae have been studied for their ability to produce EPSs. Microbial EPSs exist as homopolysaccharides or heteropolysaccharides with various properties such as different monosaccharide compositions, structural conformation, molecular weight, and functional groups. They are cost-effective alternatives to plant and animal-derived polysaccharides because the microbial cells produced them in large quantities by biotechnological processes using low-cost substrates such as industrial wastes in a short time. Microbial EPSs are safe, biodegradable, and compatible polymers. They have extensive bioactivities, including antibacterial, antifungal, antiviral, antioxidant, antitumor, antidiabetic, antiulcer, anticoagulant, antiaging, immunomodulatory, wound healing, and cholesterol-lowering activities. Microbial EPSs owing to biological activities, special biochemical structures, and attractive physicochemical properties find plenty of potential applications in various industries. The enhancement of the production of EPSs and improving their properties can be provided by genetic engineering methods. The current review aims to provide a comprehensive examination of the therapeutic activities of microbial EPSs in infectious diseases and metabolic disorders, with a focus on the mechanisms involved. Also, the effect of the physicochemical characteristics of EPSs on these bioactivities was discussed to reveal the structure-activity relationship.

The response surface methodology (RSM) was used in order to select the extraction conditions of extract from Kentichi date powder; a by-product of the date-processing process. Powder/solvent ratio, extraction temperature, and extraction time all had an impact on sugar yield, and these model factors have quadratic effects influencing sugar yield. Optimal extraction was obtained with 300 g/L powder/solvent ratio, 32.7 °C extraction temperature, and 2.1 h extraction time. Under these conditions, Kentichi date powder's (KDP) sugar yield was 77.1%, which was close to the predicted value of the model (80.50%). The results of Kentichi date powder extract (KDPE) showed that the total sugar content is 160.09 g/L. However, the protein content is 10.31 g/L with a majority of the essential amino acids (essentially glutamic acid (28.39 mg/L) and aspartic acid (9.65 mg/L)). The determination of antioxidant activity of KDPE showed a high activity (DPPH IC50 = 4.8 mg/mL, ABTS IC50 = 3 mg/mL, FRAP = 4.70 μmol AAE/mL and, TAA = 18.04 μmol Fe(II)/mL). The results show also that the freeze-drying technique has a lot of potential for producing powder from KDPE with many desirable properties. The findings indicate that KDPE with a high nutritional value could be used as a component for the formulation of functional foods.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12355-022-01223-2.

Pectinolytic enzymes that catalyze the breakdown of substrates containing pectin are widespread. Pectinases have potential applications in various industries, including food, animal feed, textile, paper, and fuel. In this study, one hundred bacterial isolates were collected from Marand city farmlands (Azarbaijan-E-Sharqi, Iran) and screened by MP medium on the base of pectinase activity considering the significance of pectinases. The results depicted that three isolates showed the most pectinase activity (more massive halo). The biochemical and molecular test results showed that the three screened bacteria were Enterobacter and named Enterobacter sp. MF41, Enterobacter sp. MF84, and Enterobacter sp. MF90. Enterobacter sp. MF84 had the largest halo, so this strain was selected for the study of its produced pectinase. The results exhibited that the produced enzyme has optimum temperature and pH for activity at 30°C and in 9, respectively. Finally, the enzyme production by Enterobacter sp. MF84 is optimized using response surface methodology (RSM) considering four factors (NH4Cl, K2HPO4, pectin, and incubation time) as variables. The results showed that the optimization procedure increased the enzyme production up to 12 times (from 1.16 to 14.16U/mg). The Pareto analysis revealed that ammonium chloride has a significant role in decreasing the enzyme production, probably by inducing the nitrification pathway enzymes in the presence of organic nitrogen in Enterobacter sp. MF84.

In recent years, there has been a growing interest in bio-based degradable plastics as an alternative to synthetic plastic. Polyhyroxybutyrate (PHB) is a macromolecule produced by bacteria as a part of their metabolism. Bacteria accumulate them as reserve materials when growing under different stress conditions. PHBs can be selected as alternatives for the production of biodegradable plastics because of their fast degradation properties when exposed to natural environmental conditions. Hence, the present study was undertaken in order to isolate the potential PHB-producing bacteria isolated from the municipal solid waste landfill site soil samples collected from the Ha’il region of Saudi Arabia to assess the production of PHB using agro-residues as a carbon source and to evaluate the growth of PHB production. In order to screen the isolates for producing PHB, a dye-based procedure was initially employed. Based on the 16S rRNA analysis of the isolates, Bacillus flexus (B. flexus) accumulated the highest amount of PHB of all the isolates. By using a UV–Vis spectrophotometer and Fourier-transform infrared spectrophotometer (FT-IR), in which a sharp absorption band at 1721.93 cm−1 (C=O stretching of ester), 1273.23 cm−1 (–CH group), multiple bands between 1000 and 1300 cm−1 (stretching of the C–O bond), 2939.53 cm−1 (–CH3 stretching), 2880.39 cm−1 (–CH2 stretching) and 3510.02 cm−1 (terminal –OH group), the extracted polymer was characterized and confirmed its structure as PHB. The highest PHB production by B. flexus was obtained after 48 h of incubation (3.9 g/L) at pH 7.0 (3.7 g/L), 35 °C (3.5 g/L) with glucose (4.1 g/L) and peptone (3.4 g/L) as carbon and nitrogen sources, respectively. As a result of the use of various cheap agricultural wastes, such as rice bran, barley bran, wheat bran, orange peel and banana peel as carbon sources, the strain was found to be capable of accumulating PHB. Using response surface methodology (RSM) for optimization of PHB synthesis using a Box–Behnken design (BBD) proved to be highly effective in increasing the polymer yield of the synthesis. With the optimum conditions obtained from RSM, PHB content can be increased by approximately 1.3-fold when compared to an unoptimized medium, resulting in a significant reduction in production costs. Thus, isolate B. flexus is a highly promising candidate for the production of industrial-size quantities of PHB from agricultural wastes and is capable of removing the environmental concerns associated with synthetic plastics from the industrial production process. Moreover, the successful production of bioplastics using a microbial culture provides a promising avenue for the large-scale production of biodegradable and renewable plastics with potential applications in various industries, including packaging, agriculture and medicine.

In the present study, the cultural conditions for exopolysaccharide (EPS) production from probiotic yeast Lipomyces starkeyi VIT-MN03 were optimized using response surface methodology (RSM) to maximize the yield of EPS. Interactions among the various factors viz. sucrose concentration (1–3 g%), NaCl concentration (2–4 g%), pH (3–5), temperature (20–30 °C), and incubation period (20–40 days) during EPS production were studied using Box-Behnken design (BBD). The EPS was purified and characterized using various instrumental analyses. The properties like adhesion, antioxidant, biosurfactant, cholesterol removal, and binding ability to mutagens were also tested for EPS produced. Sixfold increase in EPS production (4.87 g L−1) by L. starkeyi VIT-MN03 was noted under optimized condition. EPS showed a high viscosity (1.8 Pa S−1) and good shear-thinning properties. Instrumental analysis showed that EPS was heteropolysaccharide composed of glucan, mannan, and rhamnan. Lipomyces starkeyi VIT-MN03 exhibited good self-adhesion (95%) and co-aggregation ability (93%). Adhesion efficiency for yeast inoculum containing 5.5 × 107 CFU mL−1 per 9.2 cm2 of Caco-2 cell (colorectal adenocarcinoma) was noted. The probiotic EPS displayed strong antioxidant ability to scavenge hydroxyl radical and DPPH by 58% and 71% respectively. In addition, biosurfactant activity (86%) and cholesterol removal (90%) ability of probiotic EPS was also tested. EPS bound cells of L. starkeyi VIT-MN03 showed good binding ability to mutagens. These results support the effectiveness of using RSM for maximum EPS production. To the best of our knowledge, this is the first report on optimization of EPS production by probiotic yeast.

Turmeric is a spice widely used to enhance the taste and color of certain meats. We investigated the antioxidant capacities of turmeric ethanol soaking extracts by utilizing the scavenging properties of hydroxyl radical and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. The results showed that the optimum extraction parameters were: extraction time 4 h, sample-solvent ratio 1:84.12, solvent 72.34% ethanol solution, and water bath temperature 81.3 ℃. The optimum ethanol soaking extraction parameters were: extraction time 4 h, sample-solvent ratio 1:90.74, solvent 80.46% ethanol solution, water bath temperature 88.2 ℃. With these parameters, the hydroxyl radical scavenging rate and the DPPH free radical clearance of crude extracts from turmeric can reach about 93.78% and 40.69%, respectively. These results indicate that ethanol soaking extraction may be a useful method for extracting antioxidants from turmeric.

Deep eutectic solvent extraction and biological activity of polysaccharides from Tenebrio molitor

Fermented plant-based foods, including sauerkraut, offer high nutritional and functional value. Their microflora is dominated by lactic acid bacteria which are a source of different substances with health- promoting benefits and diverse applications in the food industry. Production of exopolysaccharides (EPSs) by lactic acid bacteria attracts particular interest in the food industry due to their rheological properties. In the present study, we isolated 20 strains of lactic acid bacteria from traditional Bulgarian sauerkraut. The isolates were identified by 16S rDNA sequencing and were attributed to Lactiplantibacillus plantarum (75%) and Pediococcus pentosaceus (25%). All strains were screened for their ability to synthesize exopolysaccharides, and 6 of them proved positive. Since culture media composition and especially the carbon source is a critical factor influencing the yield of bacterial EPSs, the impact of various carbon sources on the EPSs synthesis by the selected producers was explored. The best results were obtained by using glucose and sucrose as sole carbon sources.

Exopolysaccharides (EPSs) possess distinctive rheological and physicochemical properties and innovative functionality. This study aimed to investigate the physicochemical, bioactive, and rheological properties of an EPS secreted by Lactococcus lactis subsp. lactis C15. EPS-C15 was found to have an average molecular weight of 8.8 × 105 Da and was identified as a hetero-EPS composed of arabinose, xylose, mannose, and glucose with a molar ratio of 2.0:2.7:1.0:21.3, respectively. The particle size and zeta potential represented 311.2 nm and − 12.44 mV, respectively. FITR exhibited that EPS-C15 possessed a typical polysaccharide structure. NMR displayed that EPS-C15 structure is → 3)α-d-Glcvi (1 → 3)α-d-Xylv (1 → 6)α-d-Glciv(1 → 4)α-d-Glc(1 → 3)β-d-Man(1 → 2)α-d-Glci(1 → . EPS-C15 scavenged DPPH and ABTS free radicals with 50.3% and 46.4% capacities, respectively. Results show that the antiproliferative activities of EPS-C15 revealed inhibitions of 49.7% and 88.1% against MCF-7 and Caco-2 cells, respectively. EPS-C15 has antibacterial properties that inhibited Staphylococcus aureus (29.45%), Salmonella typhimurium (29.83%), Listeria monocytogenes (30.33%), and E. coli O157:H7 (33.57%). The viscosity of EPS-C15 decreased as the shear rate increased. The rheological properties of the EPS-C15 were affected by changes in pH levels and the addition of salts. EPS-C15 is a promising biomaterial that has potential applications in various industries, such as food, pharmaceuticals, and healthcare.

The significance of exopolysaccharides (EPS) in various applications has garnered increasing attention. In this study, two bacteria, Liquorilactobacillus hordei SK6 and Liquorilactobacillus mali SK26, isolated from traditional water kefir grains, produced 8.89g/L and 7.2g/L of homopolymeric glucan, respectively. NMR analysis revealed that both glucans were dextrans composed of (1 → 6)-linked α-D-glucose units, with (1 → 3)-linked α-D-glucose units serving as branching points, accounting for 5.3 ± 0.2% in dextran SK6 and 2.7 ± 0.15% in SK26. FTIR and XRD analyses further confirmed the amorphous nature of the dextrans, although dextran SK6 exhibited micro-arranged structures. Thermal characterization using TGA and DSC showed degradation temperatures of 298.5°C for dextran SK6 and 282.1°C for dextran SK26. Clear differences in morphological properties were observed using AFM and SEM. These findings provide valuable insights into dextran-producing strains and their potential applications in various industries.

Purpose: To assess exopolysaccharides (EPS) of Bacillus siamensis CV5, isolated from irradiated roots of Cistanche violacea, for their induction by ionizing radiation (IR) and their antioxidant and radioprotective power.Materials and methods: Isolated bacteria from the roots of C. violacea were screened for EPS production. The most EPS-producing bacterium was selected and the response surface methodology (RSM) was applied to elucidate the IR dose effects on EPS production. Gamma irradiation effects on the morphology and functional groups of EPS were studied using microscopy and Fourier transform infra-red (FT-IR). The radioprotective potential of EPS on the survival of B. siamensis CV5 following IR was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Free radicals scavenging potentialities (FRSP) of non-irradiated and irradiated EPS were evaluated through 2, 2−-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and ferric reducing ability of plasma (FRAP) assays.Results: Twenty strains, isolated from irradiated roots of C. violacea, were screened for their EPS production. IR-resistant B. siamensis CV5 was the most EPS-producing strain. Its purified EPS contained rhamnose, fructose, mannose and glucose. RSM indicated that EPS of CV5 (CV5-EPS) are radiation inducible. Micrographs of CV5-EPS suggested an increase in the total area and a decrease in the Feret’s statistical diameter following exposure to IR. FT-IR spectra of these EPS revealed an increase of various functional groups. The MTT survival assay demonstrated a positive correlation between the added quantity of CV5-EPS and the viability of irradiated CV5 (p < .01). DPPH, ABTS and FRAP assays indicated that the antioxidant activities of CV5-EPS increased significantly with the irradiation dose (p < .01).Conclusions: CV5-EPS were demonstrated as radiation-inducible and radioprotective biomolecules. This radioprotective potential of CV5-EPS could be associated with their antioxidant activities. In the future, irradiated EPS could be tested as a gel in cancer radiotherapy for minimizing the damage caused by rays to surrounding healthy tissues.

The exopolysaccharides (EPS) producing ability was evaluated by phenol-sulphuric acid method and the Lactobacillus HCT was the highest yield EPS-producing strain, which was isolated from feces of Bama centenarian in Guangxi of China. It was identified as Lactobacillus paracasei HCT with carbohydrate assimilation profiling and 16S rRNA gene sequencing. L. paracasei HCT was found to be a novel EPS producing strain. Preliminary one factor tests were employed to obtain the favorable conditions for EPS yield and microorganism growth in a chemically defined medium and found that carbon nitrogen ratio (C/N ratio), cultivation time and temperature had the most significant influences. Box–Behnken experimental design and response surface methodology (RSM) were adopted to further study the interactive effects of these three variables on EPS yield and cells growth. The optimal culture conditions for EPS production were: C/N ratio 9.090, cultivation time 60.67 h and temperature 29.2°C. In these conditions, the maximum EPS yield was 39.0736 mg/ml which was 4 times more than the original yield. But the optimal conditions for cell growth were: C/N ratio 8.643, 58.75 h and 31.9°C, logarithm value of viable cell count reached 7.921. These predicted values were also verified by validation experiments. Key words: Lactobacillus paracasei, exopolysaccharides, chemically defined medium, Box-Behnken design.

BackgroundCellulases are enzyme which have potential applications in various industries. Researchers are looking for potential cellulolytic bacterial strains for industrial exploitation. In this investigation, cellulase production of Bacillus cereus was explored while attacking poplar twigs. The bacterium was isolated from the gut of freshwater fish, Labeo rohita and identified by 16S rRNA gene sequencing technology. Various nutritional conditions were screened and optimized through response surface methodology. Initially, Plackett-Burman design was used for screening purpose and optimization was conducted through Box-Bhenken design.ResultsThe maximum cellulase production occurred at 0.5% yeast extract, 0.09% MgSO4, 0.04% peptone, 2% poplar waste biomass, initial medium pH of 9.0, and inoculum size of 2% v/v at 37 °C with agitation speed of 120 rpm for 24 h of submerged fermentation. The proposed model for optimization of cellulase production was found highly significant. The indigenously produced cellulase enzyme was employed for saccharification purpose at 50 °C for various time periods. Maximum total sugars of 31.42 mg/ml were released after 6 h of incubation at 50 °C.The efficiency of this enzyme was compared with commercial cellulase enzyme revealing significant findings.ConclusionThese results suggested potential utilization of this strain in biofuel industry.

The aim of this study was to optimise ultrasound-assisted extraction (UAE) of the herb Sanguisorba officinalis L. in terms of the antioxidant activity (DPPH and FRAP method) and total polyphenol content (TPC). Optimisation was performed using the response surface methodology (RSM) with a third-degree (33) Central Composite Design (CCD) approach. The RSM was applied to obtain the optimal combination of (1) raw material content (2.25–7.5 g raw material/100 mL of solvent), (2) ethanol concentration (20–60% v/v), and (3) extraction time (1–15 min). The optimal conditions for the extraction of polyphenols and antioxidant potential were a raw material content of 7.5 g/100 mL of solvent (solid/solvent ratio 13.3 mL/g), an ethanol concentration of 47% v/v, and an extraction time of 10 min. At these optimal extraction parameters, the maximum extraction of polyphenols and antioxidant activity obtained experimentally was found to be very close to its predicted value and was 12.9 mmol Trolox/L (DPPH method), 19.4 mmol FeSO4/L (FRAP method), and 2.1 g GA/L (TPC). The mathematical model developed was found to fit with the experimental data on the antioxidant potential and polyphenol extraction. The n-octanol/water partition coefficient of the optimised extract was used to determine their lipophilicity. Our studies have shown that the optimised extract is highly hydrophilic (log P &lt; 0). Optimal parameters can be used for the industrial extraction of the S. officinalis herb for the needs of, among others, the pharmaceutical or cosmetic industry.