Exopolysaccharide

Bacillus licheniformis WSF-1 strain isolated from sugar distillery plant was capable of producing a maximum amount of EPS (1.24 g/mL) and dry biomass (3.25 g/mL) respectively. Certain factors like inoculum age and carbon source was stand-ardised in view of obtaining high EPS yield. In this study, 48-hold culture was identified as the optimum inoculum age for EPS production. In fermentation, as carbon source plays a major contributing factor that influences the EPS production, optimum carbon source was determined by studying the effect of different carbohydrates such as 12% of glucose, sucrose, lactose, maltose, xylose and fructose on EPS production. The best carbon source was identified as sucrose at a concentration of 25% producing 2.9 g/mL of exopolysaccharide. The polysaccharides were hydrolysed and the monomeric components were determined as fructose and glucose by HPLC, followed by FT-IR analysis. The thermal study revealed that the EPS can withstand high temperature of 219.4 °C and had high glass transition temperature of 150.6 °C. These features indicate the thermo stable nature of the exopolysaccharide which can be used for various industrial applications. Keywords Exopolysaccharide · Bacillus · Thermo stable · Polymer · Carbon source · FTIR · SEM

Limnothrix redekei PUPCCC 116, a filamentous cyanobacterium, has been identified through 16S rRNA gene sequencing. Exopolysaccharides (EPS) of this organism have been isolated and characterized chemically, and its rheological properties were compared with commercial xanthan. The organism produced 304 µg EPS/ml culture in 21 days. The rate of EPS production was maximum (313 µg EPS/mg protein/day) during the initial days of growth, and it decreased to 140 µg EPS/mg protein/day during 18-21 days of growth. Chemical analysis of EPS revealed the presence of glucose/mannose, ribose, rhamnose, and uronic acid. Fourier transformed infrared spectrum of EPS further revealed the presence of methyl and carboxyl groups besides C–N groups indicating the presence of peptidyl moieties. Elemental analysis of EPS showed the presence of 4.97% N. The organism under continuous light produced 102% more EPS compared to when grown under a light/dark cycle of 14/10 h. The rheological properties of EPS were comparable with commercial xanthan gum.

Microbial exopolysaccharides are biothickeners that can be added to a wide variety of food products, where they serve as viscosifying, stabilizing, emulsifying or gelling agents. Numerous exopolysaccharides with different composition, size and structure are synthesized by lactic acid bacteria. The heteropolysaccharides from both mesophilic and thermophilic lactic acid bacteria have received renewed interest recently. Structural analysis combined with rheological studies revealed that there is considerable variation among the different exopolysaccharides; some of them exhibit remarkable thickening and shear-thinning properties and display high intrinsic viscosities. Hence, several slime-producing lactic acid bacterium strains and their biopolymers have interesting functional and technological properties, which may be exploited towards different products, in particular, natural fermented milks. However, information on the biosynthesis, molecular organization and fermentation conditions is rather scarce, and the kinetics of exopolysaccharide formation are poorly described. Moreover, the production of exopolysaccharides is low and often unstable, and their downstream processing is difficult. This review particularly deals with microbiological, biochemical and technological aspects of heteropolysaccharides from, and their production by, lactic acid bacteria. The chemical composition and structure, the biosynthesis, genetics and molecular organization, the nutritional and physiological aspects, the process technology, and both food additive and in situ applications (in particular in yogurt) of heterotype exopolysaccharides from lactic acid bacteria are described. Where appropriate, suggestions are made for strain improvement, enhanced productivities and advanced modification and production processes (involving enzyme and/or fermentation technology) that may contribute to the economic soundness of applications with this promising group of biomolecules.

The optimal temperature, pH and incubation time for production of exopolysaccharide (EPS) by Lactobacillus delbruckii subsp. bulgaricus and Streptococcus thermophilus strains in MRS and M17 media, respectively, were determined. In all strains, the temperature and incubation time for EPS production were 45 °C and 18 h, respectively. At 45 °C, L. delbruckiisubsp. bulgaricus B3 and G12 and S. thermophilus W22 strains produced 263, 238 and 127 mg/l, respectively. At 18 h, B3, G12 and W22 strains produced 220, 152 and 120 mg/l, respectively. While the pH for highest EPS production by L. delbruckii subsp. bulgaricus strains was 6.2 (in B3 strain: 211 mg/l, in G12 strain: 175 mg/l), for highest EPS production byS. thermophilus strain it was 6.8 (114 mg/l).

Extracellular polymeric substances (EPS) produced by a toxic dinoflagellate Amphidinium carterae Hulburt 1957 was isolated and characterized. Molecular masses of the EPS were about 233 and 1,354 kDa. Spectral analyses by 1H nuclear magnetic resonance and Fourier Transformed–Infrared Spectroscopy revealed the characteristic of the functional groups viz. primary amine, carboxyl, halide, and sulfate groups present in the EPS. However, five elements (C, O, Na, S, and Ca) were detected by scanning electron microscopy - energy dispersive X-ray spectroscopy (SEM-EDX) analysis. X-ray diffraction and differential scanning calorimetric analysis confirmed the amorphous nature of EPS, which was comprised of an average particle size of 13.969 μm (d 0.5) with 181 nm average roughness. Two monosaccharide constituents, galactose (73.13%) and glucose (26.87%) were detected by gas chromatography–mass spectroscopy analysis. Thermal gravimetric analysis revealed that degradation of EPS obtained from A. carterae takes place in three steps. The EPS produced by A. carterae was found to be beneficial for the growth of both A. carterae and Bacillus pumilus. The potential heterogeneous properties of EPS may play an important role in harmful algal bloom.

A reduced exopolysaccharide phenotype is associated with inability to synthesize polyhydroxyalkanaote (PHA) stores in Sinorhizobium meliloti strain Rm1021. Loss of function mutations in phbB and phbC result in non-mucoid colony morphology on Yeast Mannitol Agar, compared to the mucoid phenotype exhibited by the parental strain. This phenotype is attributed to reduction in succinoglycan synthesis. We have used complementation of this phenotype and the previously described d-3-hydroxybutyrate/acetoacetate utilization phenotype to isolate a heterologous clone containing a Bradyrhizobium japonicum phbC gene. Sequence analysis confirmed that this clone contains one of the five predicted phbC genes in the B. japonicum genome. The described phenotypic complementation strategy should be useful for isolation of novel PHA synthesis genes of diverse origin.

Based on the chemical composition of the exopolysaccharide produced by the cystic fibrosis bacterial isolate Burkholderia cepacia IST408, we postulated and confirmed, based on the specificity of enzymes detected in crude cell-free extracts, the pathway leading to the presumptive activated sugar precursors: UDP-D-glucose, UDP-D-galactose, UDP-D-glucuronic acid, GDP-D-mannose, and GDP-D-rhamnose. Results also suggest that regulation of the expression of the mucoid phenotype in B. cepacia does not occur at the level of synthesis of any of these enzymes.

Biofilm formation by Streptococcus pyogenes has been demonstrated as a potentially important mechanism contributing to antibiotic treatment failure. S. pyogenes is the frequent cause of purulent infections in humans and also, it could play a significant role in recurrent and chronic infections. S. pyogenes biofilm communities tend to exhibit significant tolerance to antimicrobial challenge during infections. The fluoroquinolone derivatives have been previously reported from our laboratory as effective agents against human bacterial pathogens. Therefore, in the present study, we observed the effect of these fluoroquinolones on biofilm formation. Quantitative analysis using 2,3-bis (2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide (XTT) at the biofilm inhibitory concentrations (BIC), the compounds 6a, 6c, 7b and 7c reduced 61–71% biofilm and sub-BIC (0.5 and 0.25 BIC) significantly reduced biofilm formation by up to 30–38% and 16–18%, respectively. The Fourier Transform Infrared (FTIR) spectrum of the control and treated S. pyogenes revealed the shift in the chemical entity corresponding to the exopolysaccharide (EPS). The GC/MS analysis showed that the EPS of S. pyogenes has the most abundant neutral sugars l-glucose and d-mannose which is not detected in the fluoroquinolone treated EPS.

Mutations in the Brucella melitensis quorum-sensing (QS) system are involved in the formation of clumps containing an exopolysaccharide. Here, we show that the overexpression of a gene called aiiD in B. melitensis gives rise to a similar clumping phenotype. The AiiD enzyme degrades AHL molecules and leads therefore to a QS-deficient strain. We demonstrated the presence of exopolysaccharide and DNA, two classical components of extracellular matrices, in clumps produced by this strain. We also observed that the production of outer membrane vesicles is strongly increased in the aiiD-overexpressing strain. Moreover, this strain allowed us to purify the exopolysaccharide and to obtain its composition and the first structural information on the complex exopolysaccharide produced by B. melitensis 16M, which was found to have a molecular weight of about 16 kDa and to be composed of glucosamine, glucose and mostly mannose. In addition, we found the presence of 2- and/or 6-substituted mannosyl residues, which provide the first insights into the linkages involved in this polymer. We used a classical biofilm attachment assay and an HeLa cell infection model to demonstrate that the clumping strain is more adherent to polystyrene plates and to HeLa cell surfaces than the wild-type one. Taken together, these data reinforce the evidence that B. melitensis could form biofilms in its lifecycle.

The aim of this work was to study the structure formation of yogurt made with cultures containing ropy Lactobacillus delbrueckii ssp. bulgaricus (R), capsule-forming nonropy Streptococcus thermophilus (CNR), and noncapsule-forming nonropy cultures (NCNR). Similar gelation profiles were shown for milk fermented by ropy and nonropy lactic cultures. The gelation point occurred at lower pH values in milk fermented with R or NCNR compared to that of milk fermented with CNR culture. Differences between capsule forming ropy and nonropy cultures were observed in the aggregation behavior of the caseins. Gels made with R culture had the highest maximum in loss tangent (tan delta). However, this maximum occurred at the highest pH value when CNR culture was used. The earlier gelation of milk fermented by the encapsulated nonropy strain of Strep. thermophilus resulted in increased structure rearrangement as the pH dropped, interfering with the formation of a more compact structure.

Polyhydroxyalkanoates (PHAs) is a natural, biodegradable polyester that accumulates in the form of intracellular granules in microorganisms in conditions of limiting nutrient such as nitrogen or phosphorus but in presence of excess carbon source (Doi, 1990). PHA has ...

This is the first report on the characterization and production optimization of poly-N-acetylglucosamine (PNAG) in Staphylococcus saprophyticus. A strain producing glucosamine exopolysaccharide was isolated and characterized by biochemical test and 16S rRNA gene sequence homology analysis and named as S. saprophyticus BMSZ711. The molecular mass of the purified exopolymer was about 12 kDa. Digestion of the PNAG with DispersinB proved that it has beta-1,6 linkage. BMSZ711 can only produce PNAG when grown in M1 minimal medium but not in nutrient rich medium with optimum temperature of 30 degrees C and pH of 7. Glycerol and ammonium sulfate were found to be the best carbon and nitrogen source, respectively. Maximum PNAG production was obtained when glycerol 100mM, ammonium sulfate 0.3%, yeast extract 1.5 g/L, sodium chloride 10 g/L and valine 2mM were used.