Monosaccharide Composition Of Exopolysaccharides Research Articles

Quantitative image analysis of influence of polysaccharides on protein network formation in GDL-acidified milk gels

Exopolysaccharides (EPS) are commonly used to improve the texture of yogurt. These polysaccharides interact with casein micelles, the major protein in milk, via electrostatic and depletion mechanisms during fermentation by lactic acid bacteria (LAB). However, the relationship between the physicochemical properties and monosaccharide composition of EPS and their impact on yogurt texture is not yet fully understood. To address this knowledge gap, we studied the effects of polysaccharides commonly used as food additives on acid-induced milk protein networks. Confocal laser scanning microscopy (CLSM) was used to image the network microstructures. Image analysis, including Fourier transform, autocorrelation, and binarization-based techniques, was applied to quantify key structural features of the mixed milk protein/polysaccharide gels. These parameters were then related to the macroscopic properties of the model food matrices, such as elastic and viscous moduli and yield point. We found that the addition of neutral polysaccharides resulted in a concentration-dependent increase in structure factor, protein domain size, and pore fraction. In contrast, the presence of charged polysaccharides led to an increase in protein domain size, a decrease in pore fraction, and a decrease in elastic and viscous moduli. These results demonstrate the use of a quantitative image analysis method for selecting LAB with favorable EPS properties to improve yogurt texture.

Inulin increases the EPS biosynthesis of Lactobacillus delbrueckii ssp. bulgaricus LDB-C1.

Its eps gene cluster, the antioxidant activity and monosaccharide composition of exopolysaccharides, the expression levels of related genes at different fermentations were analyzed for clarifying the exopolysaccharide biosynthesis mechanism of Lactobacillus delbrueckii subsp. bulgaricus LDB-C1. The comparison analysis of eps gene clusters indicated that the gene clusters present diversity and strain specificity. The crude exopolysaccharides from LDB-C1 exhibited a good antioxidant activity. Compared with glucose, fructose, galactose, and fructooligosaccharide, inulin significantly improved the exopolysaccharide biosynthesis. The structures of EPSs were significantly different under different carbohydrate fermentation conditions. Inulin obviously increased the expressions of most EPS biosynthesis related genes at fermentation 4h. Inulin accelerated the beginning of the exopolysaccharide production in LDB-C1, and the enzymes promoted by inulin was beneficial for the accumulation of exopolysaccharide at the whole fermentation process.

Sustainable use of agro-industrial wastes as potential feedstocks for exopolysaccharide production by selected Halomonas strains.

Large quantities of waste biomass are generated annually worldwide by many industries and are vastly underutilized. However, these wastes contain sugars and other dissolved organic matter and therefore can be exploited to produce microbial biopolymers. In this study, four selected Halomonas strains, namely, Halomonas caseinilytica K1, Halomonas elongata K4, Halomonas smyrnensis S3, and Halomonas halophila S4, were investigated for the production of exopolysaccharides (EPS) using low-cost agro-industrial wastes as the sole carbon source: cheese whey, grape pomace, and glycerol. Interestingly, both yield and monosaccharide composition of EPS were affected by the carbon source. Glucose, mannose, galactose, and rhamnose were the predominant monomers, but their relative molar ratio was different. Similarly, the average molecular weight of the synthesized EPS was affected, ranging from 54.5 to 4480kDa. The highest EPS concentration (446mg/L) was obtained for H. caseinilytica K1 grown on cheese whey that produced an EPS composed mostly of galactose, rhamnose, glucose, and mannose, with lower contents of galacturonic acid, ribose, and arabinose and with a molecular weight of 54.5kDa. Henceforth, the ability of Halomonas strains to use cost-effective substrates, especially cheese whey, is a promising approach for the production of EPS with distinct physicochemical properties suitable for various applications.

Expression of genes involved in exopolysaccharide synthesis in Lactiplantibacillus plantarum VAL6 under environmental stresses.

Environmental factors can alter exopolysaccharide biosynthesis in lactic acid bacteria (LAB). To further clarify this potential relationship, the mRNA expression of genes involved in exopolysaccharide synthesis such as glmU, pgmB1, cps4E, cps4F, cps4J, and cps4H in Lactiplantibacillus plantarum VAL6 under different conditions including temperature, pH, sodium chloride (NaCl), and carbon dioxide (CO2) intensification culture was studied. The transcriptomic data revealed that the exposure of L. plantarum VAL6 at pH 3 increased the expression level of cps4H but decreased the expression levels of pgmB1 and cps4E. Under pH 8, cps4F and cps4E were significantly upregulated, whereas pgmB1 was downregulated. Similarly, the expression levels of cps4F, cps4E, and cps4J increased sharply under stresses at 42 or 47°C. In the case of NaCl stress, glmU, pgmB1, cps4J, and cps4H were downregulated in exposure to NaCl at 7 and 10% concentrations while cps4E and cps4F were upregulated at 1h of 10%-NaCl treatment and at 5h of 4%-NaCl treatment. Remarkably, CO2 intensification culture stimulated the expression of all tested genes. In addition, simultaneous changes in expression of cps4E and cps4F under environmental challenges may elicit the possibility of an association between the two genes. These findings indicated that the expression level of eps genes is responsible for changes in the yield and monosaccharide composition of exopolysaccharides under environmental stresses.

The effect of optimized carbon source on the synthesis and composition of exopolysaccharides produced by Lactobacillus paracasei

This study aimed to predict the optimal carbon source for higher production of exopolysaccharides (EPS) by Lactobacillus paracasei TD 062, and to evaluate the effect of this carbon source on the production and monosaccharide composition of EPS. We evaluated the EPS production capacity of 20 strains of L. paracasei under the same conditions. We further investigated L. paracasei TD 062, which showed the highest EPS-producing activity (0.609 g/L), by examining the associated biosynthesis pathways for EPS. Genomics revealed that fructose, mannose, trehalose, glucose, galactose, and lactose were carbon sources that L. paracasei TD 062 could use to produce EPS. We identified an EPS synthesis gene cluster that could participate in transport, export, and sugar chain synthesis, and generate 6 sugar nucleotides. Experimental results showed that the sugar content of the EPS produced using fermentation with the optimized carbon source (fructose, mannose, trehalose, glucose, galactose, and lactose) increased by 115%. Furthermore, use of the optimized carbon source changed the monosaccharide content of the associated EPS. The results of enzyme activity measurements showed significant increases in the activity of 2 key enzymes involved in the glycoside synthesis pathway. Our study revealed that optimizing the carbon source provided for fermentation not only increased the production of EPS, but also affected the composition of the monosaccharides by increasing enzyme activity in the underlying synthesis pathways, suggesting an important role for carbon source in the production of EPS by L. paracasei TD 062.

Response of Lactobacillus plantarum VAL6 to challenges of pH and sodium chloride stresses

To investigate the effect of environmental stresses on the exopolysaccharide biosynthesis, after 24 h of culture at 37 °C with pH 6.8 and without sodium chloride, Lactobacillus plantarum VAL6 was exposed to different stress conditions, including pH (pHs of 3 and 8) and high sodium chloride concentration treatments. The results found that Lactobacillus plantarum VAL6 exposed to stress at pH 3 for 3 h gives the highest exopolysaccharide yield (50.44 g/L) which is 6.4 fold higher than non-stress. Under pH and sodium chloride stresses, the mannose content in exopolysaccharides decreased while the glucose increased in comparison with non-stress condition. The galactose content was highest under stress condition of pH 8 meantime rhamnose content increased sharply when Lactobacillus plantarum VAL6 was stressed at pH 3. The arabinose content in exopolysaccharides was not detected under non-stress condition but it was recorded in great amounts after 3 h of stress at pH 3. In addition, stress of pH 8 triggered the mRNA expression of epsF gene resulting in galactose-rich EPS synthesis. According to our results, the stresses of pH and sodium chloride enhance the production and change the mRNA expression of epsF gene, leading to differences in the monosaccharide composition of exopolysaccharides.

Optimization of production and intrinsic viscosity of an exopolysaccharide from a high yielding Virgibacillus salarius BM02: Study of its potential antioxidant, emulsifying properties and application in the mixotrophic cultivation of Spirulina platensis

Virgibacillus salarius BM02 was identified as a highly exopolysaccharide (EPS) producing bacterium. The EPS production and its physico-chemical properties (intrinsic viscosity and total sugars/protein (TS/P) ratio) were optimized using Box-Behnken experimental design. Maximum EPS production of 5.87 g L−1 with TS/P ratio of 12.56 and intrinsic viscosity of 0.13 dL g−1 was obtained at optimal conditions of sucrose (4.0% w/v), peptone (0.75% w/v) and incubation period of 4.69 day. The monosaccharide composition of EPS was mannose, arabinose and glucose at a molar ratio of 1.0:0.26:0.08. The EPS showed high water solubility (38.5%), water holding capacity (514.46%) and foaming capacity (55.55%). The EPS showed moderate antioxidant activity in vitro and good emulsion stabilizing properties against several hydrophobic compounds. The emulsifying activity was stable at different temperatures, pH and ionic strength. Additionally, the acid hydrolysate of the EPS was evaluated as a carbon source for the mixotrophic cultivation of industrially important Spirulina platensis. It induced an enhancement of not only biomass production of S. platensis, but also cellular contents (pigments, proteins and lipids) leading to higher nutritional value.

Production, structural characterization, and antiproliferative activity of exopolysaccharide produced by Scleroderma areolatum Ehrenb with different carbon source.

The effects of different three carbon sources, that is, glucose, fructose, and sucrose, on production, molecular properties and antiproliferative activity of exopolysaccharide (EPS), were evaluated in the submerged culture of Scleroderma areolatum Ehrenb. Among carbon sources examined, the addition of sucrose maximizes the mycelia production, while fructose could maximize the EPS yield. Although the predominant carbohydrate compositions identified were gluconic acid and mannose, the monosaccharide composition of EPSs was also different significantly. FT-IR spectral analysis revealed there was no significant difference among the prominent characteristic groups in three EPSs. The molecular weight of EPSs was also affected by carbon source, being generally lower compared with that with glucose. However, all EPSs molecule existed as nearly globular shape form in aqueous solution. The variation of carbon sources also affected antiproliferative activity examined in vitro using cell proliferation assay. Fructose was optimal carbon source giving higher antiproliferative activity probably due to the relatively high contents of xylose in the EPS with low molecular weight.

Characterization of highly branched dextran produced by Leuconostoc citreum B-2 from pineapple fermented product

A strain Leuconostoc citreum B-2 was isolated from homemade fermentation product of pineapple and its polysaccharide yield was 28.3g/L after cultivating the strain in Man-Rogosa-Sharpe (MRS) medium with 75g/L sucrose. The exopolysaccharide (EPS) was characterized by gas chromatography (GC), Fourier-transform infrared (FT-IR) spectra, high-performance size-exclusion chromatography (HPSEC), nuclear magnetic resonance (NMR) spectroscopy and scanning electron microscope (SEM) analysis in present study. The monosaccharide composition of EPS was glucose and molecular weight was 3.77×106Da. FT-IR and NMR spectra revealed that the B-2 EPS was composed of 75% α-(1→6) linked d-glucopyranose units existing in the main chain with 19% α-(1→3) branching and only a few α-(1→2) branching. The SEM of the dried EPS appeared irregular sheets with glittering surface and compact structure. Water solubility index and water holding capacity of B-2 EPS were 80% and 450%, respectively. All the mentioned characteristics suggested that the EPS has a potential application in the food, cosmetic and pharmaceuticals industry.

Characterization of the chemical structures and physical properties of exopolysaccharides produced by various Streptococcus thermophilus strains

Exopolysaccharides (EPS) produced by some lactic acid bacteria are often used by the dairy industry to improve the rheological and physical properties of yogurt, but the relationship between their structure and functional effect is still unclear. The EPS from different species, or different strains from the same species, may differ in terms of molar mass, repeating unit structure, and EPS yield during fermentation of milk. This study aimed to characterize the detailed properties of EPS produced from 7 strains of Streptococcus thermophilus, which is one of the key cultures used for yogurt manufacture. Milk was fermented with strains DGCC 7698, DGCC 7710, DGCC 7785, ST-10255y, St-143, STCth-9204, and ST4239. These strains were selected because they have been used in previous studies on yogurt texture, but a complete description of their EPS structural properties has not yet been reported. All strains were fermented under a similar acidification rate by adjusting the level of supplementation with peptone or the inoculation level, which allowed for a comparison of EPS yields under similar growth conditions (reconstituted skim milk at 40°C). The EPS from each strain was isolated and the weight-average molar mass and z-average root mean square radius determined using size-exclusion chromatography multiangle laser light scattering. The monosaccharide composition of EPS was determined using gas chromatography-mass spectrometry, and repeating unit structure was determined using nuclear magnetic resonance spectroscopy. The weight-average molar mass values of EPS ranged from 0.14 to 1.61 × 106 g/mol. All 7 EPS samples were uncharged. The strains ST-10255y and ST4239 had EPS with the same repeating unit structure. The monosaccharide compositions of the various EPS were mainly composed of glucose and galactose, with low levels of rhamnose in the EPS isolated from DGCC 7698, and N-acetylgalactosamine in the EPS from DGCC 7785, ST-10255y, and ST4239. The yields of EPS (measured when fermented milks reached pH 4.6) ranged from 8.0 to 76.4 mg of glucose equivalents/kg. In addition to (free) EPS, some strains were also able to produce capsular polysaccharide (associated with the bacterial cells) when observed with negative staining technique. The results of our study will help the dairy industry to better understand the mechanism by which different strains of Streptococcus thermophilus affect yogurt texture.

Effects of mixed carbon sources on galactose and mannose content of exopolysaccharides and related enzyme activities in Ganoderma lucidum

This work investigated the effects of mixed carbon sources on the monosaccharide composition of exopolysaccharides (EPS) and activity of related enzymes, phosphoglucose isomerase (PGI), α-phosphoglucomutase (PGM), phosphomannose isomerase (PMI) and GDP-D-Man pyrophosphorylase (GMP), in the EPS biosynthesis by Ganoderma lucidum. Combinations of two carbon sources, glucose (Glc) + mannose (Man) and Glc + galactose (Gal), were applied at various mass ratios in the liquid culture of G. lucidum. The results showed that Glc, Gal, and Man were the major monosaccharides in G. lucidum EPS, and the combined mole percentages of monosaccharides were >80%, the same as those in the carbon source mixture. Activities of PGI, PMI, and GMP were correlated with Man mole percentage in EPSs and were enhanced by increasing the proportion of Gal for the carbon source Glc + Gal. PGM activity was correlated with Gal mole percentage and was reduced by increasing the Man proportion for Glc + Man. The expression of four genes encoding PGI, PMI, GMP and PGM reached peak levels on day 6 of culture, whereas the enzyme activities increased steadily from day 3 to 8. The monosaccharide composition of G. lucidum EPS for various mixed carbon source conditions thus appeared to be controlled by the translational level of genes encoding PGM, PGI, PMI, and GMP. These findings will be helpful to control the monosaccharide composition for desired biological activity of EPS produced by G. lucidum fermentation.

Production and characterization of exopolysaccharides in mycelial culture ofCordyceps sinensisfungus Cs-HK1 with different carbon sources

The effects of different carbon sources (sugars) on the production and molecular properties of exopolysaccharides (EPS) were evaluated in the mycelial liquid culture of a medicinal fungusCordyceps sinensisCs-HK1. Galactose or mannose was used (at 5g·L−1) as a secondary carbon source with glucose (35g·L−1) at the mass ratio of 1:7. Mannose was consumed notably since the first day of culture, but galactose was not even after glucose was exhausted. The volumetric yield of EPS in culture was increased slightly with the addition of galactose and decreased with mannose. The monosaccharide composition of EPS was also different,e.g., on day 8, the glucose contents of EPS were 76% with the addition of mannose, 59% with galactose, compared with 62% with glucose only. The molecular weight distribution of EPS was also affected by the secondary carbon source, being generally lower compared with that with glucose only. The results suggested that the addition of galactose improved the total yield of EPS in culture while mannose can improve the yield of glucan constituent of EPS.

Diatom Exopolysaccharides: a Review

Diatom exopolysaccharides (EPS) are bioactive components released into the environment. As an important component of marine phytoplankton, diatoms produce up to a quarter of the annual primary organic matter. The results of EPS studies highlighting the important role of diatom EPS in the global carbon cycle are analyzed. The main types of extracellular carbohydrates, monosaccharide composition of EPS, and dependence of their production on algal growth rate, light conditions and nutrient content in the environment are discussed.

Assessment of the capability of a gelling complex made of tara gum and the exopolysaccharides produced by the microorganism Streptococcus thermophilus ST10 to prospectively restore the gut physiological barrier: a pilot study.

Leaky gut, or intestinal permeability, is the phenomenon of the gut wall exhibiting increased absorbency. It is pretty well recognised that an altered or damaged bowel lining or gut wall may result from unbalanced diet, parasites, infection, or medications and that this allows substances such as toxins, microbes, undigested food, or waste to leak through. As a natural consequence, this prompts the body to initiate an immune reaction leading to potentially severe health conditions. Different strategies may be used to improve, at least temporarily, the physiological intestinal barrier. The use of specific beneficial microorganisms, such as lactobacilli and bifidobacteria, has been suggested as an innovative tool to counteract an improper level of intestinal permeability. The association of bacteria with specific gelling agents, such as gums, may represent an improvement since these molecules are able to form hydrophilic gels that distribute uniformly over the inner intestinal surface. This pilot study was undertaken to evaluate intestinal permeability in subjects treated with a gelling complex, an association of tara gum and the microorganism Streptococcus thermophilus ST10 (DSM 25246), which has a well-demonstrated in vitro ability to synthesise and secrete exopolysaccharides (EPSs). Twenty-five healthy subjects were enrolled in this human intervention, double-blind, placebo-controlled, pilot trial (age between 21 and 57 y, mean 37.7±11.2). Subjects were then randomised into 2 groups: group A (13 subjects) was given an active formulation containing 250 mg of tara gum and 1 billion viable cells of S. thermophilus ST10, whereas group B (12 subjects) was given a placebo formulation. All the subjects participating in the study were directed to take 1 dose per day for 30 consecutive days. The presence and concentration of exopolysaccharides (EPSs) in the faeces was determined at time 0 (d0), after 30 days of treatment (d30), and at the end of the 2-week follow-up period (d45). The monosaccharide composition of EPSs was used to quantify the possible contribution of tara gum to the amount of polysaccharides detected in the faecal material. Intestinal permeability was evaluated at the same time by means of the lactitol/mannitol ratio (small intestine permeability) and sucralose concentration (colonic permeability) in urine specimens sampled after specified times. A statistical comparison was made between the concentration of EPSs, the lactulose/mannitol ratio, and the amount of excreted sucralose in the 2 groups at d0, d30, and d45. In the active group, supplementation with S. thermophilus ST10 and tara gum was able to significantly increase the faecal EPSs concentration compared with placebo (from 0.169 mg/g to 0.633 mg/g after 30 d, P<0.001). An interesting decrease in intestinal permeability, both of the small bowel and in the colon, was also recorded. The L/M ratio diminished from 0.021 in the active group to 0.014 and 0.015 after 30 and 45 days, respectively (P=0.045 and P=0.033 compared with placebo). The sucralose concentration decreased from 35.8 mg to 27.9 mg and 29.1 mg (P=0.038 and P=0.026 compared with placebo) at the end of the supplementation period and after the follow-up, respectively. No significant differences were recorded in the placebo after 30 days or at the end of the follow-up. The association of the EPSs produced by S. thermophilus ST10 and tara gum seems capable of significantly improving the intestinal functional barrier in healthy subjects. A wider study in subjects presenting impaired gut permeability would be useful in the future to confirm the positive results from this pilot trial. In any case, our findings are consistent with the parallel increase in exopolysaccharide concentration in the faecal material, thus suggesting the effective ability of the strain used to secrete EPSs in the gut lumen. An innovative approach of this type may be useful in helping to restore the physiological barrier by means of a merely natural and mechanical action.

Influences of carbon sources on the biomass, production and compositions of exopolysaccharides from Paecilomyces hepiali HN1

The parasitic fungus, Paecilomyces hepiali, is used to produce Cordyceps materials as succedaneum of natural Cordyceps sinensis in China. The purpose of this research was to investigate the effects of glucose, mannose, sucrose, lactose as solo carbon source and sucrose + lactose or mannose + sucrose as synthetic carbon source on the growth of mycelium and production, chemical composition, molecular weight distribution and monosaccharide composition of exopolysaccharides from P. hepiali HN1 (PHEPS). The maximum mycelium biomass of 12.16 kg m−3 and PHEPS yield of 4.57 kg m−3 were achieved from the culture with sucrose (50 kg m−3) as carbon source. The resulting PHEPS was characterized by analyses of chemical composition, size-exclusion chromatography and high performance liquid chromatography with 1-phenyl-3-methyl-5-pyrazolone pre-column derivatization. It was found that the chemical compositions and monosaccharide ratios in PHEPS were significantly affected by the carbon sources used. Glucose or mannose as carbon source enhanced the biosynthesis of PHEPS with higher-molecular weight (>1000 kD), but solo carbon source of lactose or synthetic carbon source of mannose + lactose did not increase the ratio of galactose in PHEPS. The metabolism kinetics of carbon sources demonstrated the correlation between PHEPS synthesis and the utilization of carbon sources. These findings will be useful for further works on the production, structure and function of PHEPS.

Identification of α-d-glucose-1-phosphate cytidylyltransferase involved in Ebosin biosynthesis of Streptomyces sp. 139

Ebosin, a novel exopolysaccharide produced by Streptomyces sp. 139 has antagonist activity for IL-1R in vitro and remarkable anti-rheumatic arthritis activity in vivo. Its biosynthesis gene cluster (ste) has been identified. In this study, gene ste17 was expressed in Escherichia coli BL21 and the recombinant protein was purified. With CTP and alpha-D-glucose-1-phosphate as substrates, the recombinant Ste17 protein was found capable of catalyzing the production of CDP-D-glucose and pyrophosphate, demonstrating its identity as an alpha-D-glucose-1-phosphate-cytidylyltransferase (CDP-D-glucose synthase). To investigate the function of ste17 in Ebosin biosynthesis, the gene was disrupted with a double crossover via homologous recombination. The monosaccharide composition of exopolysaccharide (EPS) produced by the mutant Streptomyces sp. 139 (ste17 (-)) was found significantly altered from that of Ebosin, with glucose becoming undetectable. This gene knockout also negatively affected the antagonist activity for IL-1R of EPS. These results indicate that the CDP-D: -glucose synthase encoded by ste17 gene is involved in the formation of nucleotide sugar (CDP-D-glucose) as glucose precursor in Ebosin biosynthesis.

Benthic mucilaginous aggregates in the Mediterranean Sea: Origin, chemical composition and polysaccharide characterization

Benthic mucilaginous aggregates are frequently formed in various parts of the Mediterranean basin, as in the Tyrrhenian and Adriatic Seas. Notwithstanding their wide spatial distribution, the role played by these aggregates in the biogeochemical cycling of organic matter is still largely unknown. The characteristics of the benthic aggregates examined in the present study showed that they are different from other mucilaginous aggregates, such as the “pelagic” ones that can form in the same areas in the water column and subsequently sediment to the seafloor. The aggregates are usually formed of structures of the filamentous macroalgae Acinetospora crinita, Chrysonephos lewisii and Nematochrysopsis marina. The elemental composition of the aggregates showed a marine macrophyte origin on the basis of the bulk organic matter content of the aggregates. Carbohydrates and proteins account for 26.6 to 55.9% of the organic carbon in the mucilage, respectively. Monosaccharide composition of exopolysaccharides in the mucilage aggregates revealed a characteristic pattern, with galactose, xylose or mannose and fucose as the major components. The relatively high content of deoxysugars is another distinctive feature. The abundant sulphate and uronic groups present in the polysaccharides in addition to their macromolecular dimensions and elongation contribute to inter-chain aggregation. Electron microscopic observations suggest that the polysaccharide fraction is the main macromolecular component in the formation of the persistent gel network in the aggregates.