Optimization and characterization of an extracellular polysaccharide produced byMoniliella pollinis

Summary A yeast strainMoniliellapollinis produces an extracellular highly viscous gum-like polysaccharide of glucan type in a simple mineral medium. Optimum conditions for its production and properties are described. The viscosity decreased after lyophilization.     

Optimization and characterization of an extracellular polysaccharide produced byGlomerella cingulata

Summary A new strain of the fungusGlomerellacingulata has been isolated, which produces an extracellular highly viscous polysaccharide in a simple mineral medium. Optimum conditions for its production and properties are described. The polysaccharide produced was a glucan type. The viscosity remained stable during storage over a period of seven days. Large changes in temperature and pH have no effect on viscosity.     

Optimization, purification, characterization and antioxidant activity of an extracellular polysaccharide produced by Paenibacillus polymyxa SQR-21

The optimization, purification and characterization of an extracellular polysaccharide (EPS) from a bacterium Paenibacillus polymyxa SQR-21 (SQR-21) were investigated. The results showed that SQR-21 produced one kind of EPS having molecular weight of 8.96 × 10⁵ Da. The EPS was comprised of mannose, galactose and glucose in a ratio of 1.23:1.14:1. The ratio of monosaccharides and glucuronic acid was 7.5:1. The preferable culture conditions for EPS production were pH 6.5, temperature 30 °C for 96 h with yeast extract and galactose as best N and C sources, respectively. The maximum EPS production (3.44 g L⁻¹) was achieved with galactose 48.5 g L⁻¹, Fe³⁺ 242 μM and Ca²⁺ 441 μM. In addition, the EPS showed good superoxide scavenging, flocculating and metal chelating activities while moderate inhibition of lipid peroxidation and reducing activities were determined. These results showed the great potential of EPS produced by SQR-21 to be used in industry in place of synthetic compounds.     

Partial characterization of an extracellular polysaccharide produced by the moderately halophilic bacterium Halomonas xianhensis SUR308

A moderately halophilic bacterium, Halomonas xianhensis SUR308 (Genbank Accession No. KJ933394) was isolated from a multi-pond solar saltern at Surala, Ganjam district, Odisha, India. The isolate produced a significant amount (7.87 g l^?1) of extracellular polysaccharides (EPS) when grown in malt extract–yeast extract medium supplemented with 2.5% NaCl, 0.5% casein hydrolysate and 3% glucose. The EPS was isolated and purified following the conventional method of precipitation and dialysis. Chromatographic analysis (paper, GC and GC-MS) of the hydrolyzed EPS confirmed its heteropolymeric nature and showed that it is composed mainly of glucose (45.74 mol%), galactose (33.67 mol %) and mannose (17.83 mol%). Fourier-transform infrared spectroscopy indicated the presence of methylene and carboxyl groups as characteristic functional groups. In addition, its proton nuclear magnetic resonance spectrum revealed functional groups specific for extracellular polysaccharides. X-ray diffraction analysis revealed the amorphous nature (CI_xrd, 0.56) of the EPS. It was thermostable up to 250°C and displayed pseudoplastic rheology and remarkable stability against pH and salts. These unique properties of the EPS produced by H. xianhensis indicate its potential to act as an agent for detoxification, emulsification and diverse biological activities.     

Preparation,structural characterization and antioxidant activity of an extracellular polysaccharide produced by the fungus Oidiodendron truncatum GW

A water-soluble extracellular polysaccharide, designated Os2-1, was isolated from the fermented liquid of the fungus Oidiodendron truncatum GW using ethanol precipitation, anion-exchange and gel-permeation chromatography. Os2-1 was mainly composed of glucose with minor amounts of glucosamine, and its average molecular weight was about 9.6 kDa. On the basis of chemical and spectroscopic analyses, including one- and two-dimensional nuclear magnetic resonance (1D and 2D NMR) spectroscopy, the backbone of Os2-1 was characterized to mainly consist of (1→6)-linked α-d-glucopyranose residues with minor amounts of (1→2)-linked α-d-glucopyranose residues. The antioxidant activity of Os2-1 was evaluated with the scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide radicals and inhibition effect of lipid peroxidation in vitro, and the results indicated that Os2-1 had good antioxidant activity, especially scavenging ability on DPPH and superoxide radicals. The investigation demonstrated that Os2-1was an extracellular polysaccharide differing from previously described extracellular polysaccharides, and could be a potential antioxidant.     

Isolation of an extracellular polysaccharide (EPS) depolymerase produced byBradyrhizobium japonicum

Bradyrhizobium japonicum is capable of producing an acidic, high molecular weight, extracellular polysaccharide (EPS). An enzyme exhibiting EPS depolymerase activity was detected in cell lysates ofB. japonicum strain 2143. The depolymerase was active against the EPS produced by strain 2143 and the closely related EPS produced by strain 311b 110. Depolymerase activity was characterized by its ability to decrease the viscosity of EPS solutions, to decrease the molecular weight of EPS, and to catalyze the release of reducing groups from EPS. The depolymerase exhibited a sharp activity optimum at pH 6 and had a molecular weight of approximately 45 kD as determined by gel permeation chromatography. Analysis of depolymerase-treated EPS indicates that the enzyme acts as an endo-depolymerase, producing a relatively narrow size range of high molecular weight products.Contribution from the Missouri Agricultural Experiment Station, Journal Series Number 10:959.     

Isolation and characterization of a new extracellular polysaccharide from an Acetobacter species

An expolysaccharide (EPS)-producing strain of the genus Acetobacter (named IFR 101) was isolated from a commercial vinegar acidifier. IFR 101 can be grown over the pH range 3.0–8.0 and the temperature range 12–35°C, and is ethanol-tolerant to 10% v/v. Neutral sugar analysis of the purified EPS indicates the presence of the sugars mannose, galactose and glucose. Methylation studies and NMR data suggest that the EPS may either be a complex branched polysaccharide or a mixture of simpler linear and branched structures. Aqueous preparations of IFR 101 exhibit reversible shear-thinning behaviour.     

An extracellular polysaccharide produced by Zoogloea ramigera 115

A weakly acidic polysaccharide was purified from the extracellular zoogloeal matrix produced by Zoogloeal ramigera 115. The purified polysaccharide was homogeneous as judged by sedimentation analysis, and the average molecular weight was estimated to be about 10(5) by gel permeation chromatography of the fully methylated preparation. The polysaccharide was composed of D-glucose, D-galactose and pyruvic acid in an approximate molar ratio 11:3:1.5. On the basis of methylation, periodate oxidation, Smith degradation and partial hydrolysis, the following highly branched structure was deduced for the polysaccharide: a long chain mainly consisting of beta 1 leads to 4-linked glucose residues branching at the C-3 or C-6 position of galactose residues which are present in beta 1 leads to 4 or beta 1 leads to 3 linkages as the minor component of the long chain; pyruvic acid residues, the sole acidic component, are linked to the nonreducing end and/or 1,3-linked glucose residues through 4,6-ketal linkages. The purified polysaccharide was not readily soluble in water and had a high affinity for several metallic ions (e.g, 0.25 mumol Fe3+/mg, and 0.17 mumol Fe2+ mg). Upon addition of metallic ions (1 mM) to a gelatinous aqueous solution of the polysaccharide (K+ form, 0.125%), more than 80% of it immediately coprecipitated out with them.     

Physical properties of an extracellular polysaccharide produced by Bacillus sp. CP912

AIMS: In this study, some physical properties of Bacillus sp. exo-polysaccharide were investigated. METHODS AND RESULTS: An extracellular polysaccharide was purified by sequential precipitations after homogenization of the diluted culture supernatant of Bacillus sp. CP912. Its physical properties were examined such as lipid emulsifying effect on several vegetable oils and flocculating activity against the activated carbon suspension. The melting point and endothermic calories of the polysaccharide were 128.7 degrees C and 50.864 kCal mol-1, respectively. Its pyrolysis temperature was 284.58 degrees C. The polysaccharide showed high lipid emulsifying activity on oil-water emulsion, against olive, peanut, sunflower and corn oils. It exhibited high flocculating activity as well against activated carbon. CONCLUSIONS: The present findings suggest that the extracellular polysaccharide produced by Bacillus sp. CP912 has a great industrial potential because of its high lipid emulsifying and flocculating activity. SIGNIFICANCE AND IMPACT OF THE STUDY: These data represent a novel Bacillus sp. extracellular polysaccharide possessing high emulsifying and flocculating effects.     

Characterization of an extracellular polysaccharide produced by a Pseudomonas strain grown on glycerol

A new extracellular charged polysaccharide composed mainly by galactose, with lower amounts of mannose, glucose and rhamnose, was produced by the cultivation of Pseudomonas oleovorans NRRL B-14682 using glycerol as the sole carbon source. Thermal and solid-state NMR analysis showed that this polymer is essentially amorphous, with a glass transition temperature of 155.7 degrees C. The exopolysaccharide aqueous solutions have viscoelastic properties similar to that of Guar gum, but with affinity to salts as a result of its polyelectrolyte character. In addition, the exopolysaccharide has demonstrated good flocculating and emulsifying properties and film-forming capacity. These properties make this polymer a good alternative to more expensive natural polysaccharides, such as Guar gum, in several applications in the food, pharmaceutical, cosmetic, textile, paper and petroleum industries.     

Structural analysis of an extracellular polysaccharide produced by Rhodococcus rhodochrous strain S-2

A possibility has been suggested of applying the EPS produced by Rhodococcus rhodochrous strain S-2 (S-2 EPS) to the bioremediation of oil-contaminated environments, because its addition, together with minerals, to oil-contaminated seawater resulted in emulsification of the oil, increased the degradation of polyaromatic hydrocarbons (PAH) of the oil, and led to the dominance of PAH-degrading marine bacteria. To understand the underlying principles of these phenomena, we determined the chemical structure of the sugar chain of S-2 EPS. The EPS was found to be composed of D-galactose, D-mannose, D-glucose, and D-glucuronic acid, in a molar ratio of 1:1:1:1. In addition, 0.8% (w/w) of octadecanoic acid and 2.7% (w/w) of hexadecanoic acid were also contained in its structure. By 1H and 13C NMR spectroscopy, including 2D DQF-COSY, TOCSY, HMQC, HMBC, and NOESY experiments, as well as chemical and enzymatic analyses, the polysaccharide was shown to consist of tetrasaccharide repeating units with the following structure: (see formula in text).     

Purification and characterization of an extracellular beta-lactamase produced by Acinetobacter calcoaceticus.

A beta-lactamase was purified 430-fold from the culture supernatant of Acinetobacter calcoaceticus by ion exchange chromatography on CM-Sephadex and affinity chromatography on phenylboronic-acid-agarose. The purified enzyme was homogeneous as judged by SDS-PAGE, and was characterized with respect to molecular mass (38 and 41 kDa by gel filtration on Sephadex G-75 and SDS-PAGE, respectively), pH optimum (pH 7.0), temperature optimum (45 degrees C) and isoelectric point (9.3). The beta-lactamase showed mainly cephalosporinase activity. It was inhibited by cloxacillin, carbenicillin, penicillanic acid sulphone (sulbactam) and aztreonam. It was not inhibited by clavulanic acid up to a concentration of 0.25 mM. Neither EDTA nor p-chlormercuribenzoate, up to concentrations of 1 or 100 mM, respectively, affected activity. According to these characteristics, it is a typical CEP-N cephalosporinase.     

Optimization and characterization of a polysaccharide produced by Pseudomonas fluorescens WR-1 and its antioxidant activity

The extracellular polysaccharide produced by a newly isolated strain Pseudomonas fluorescens WR-1 was purified and characterized and its production was optimized using response surface methodology. The results showed that the strain WR-1 produced one kind of EPS that was composed of arabinose, glucose and uronic acid. The molecular weight of the EPS was determined to be 6.78×10(6)Da. The preferable culture conditions for EPS production were pH 7.0, temperature 28°C for 72h with peptone and maltose as best N and C sources, respectively. The model predicted that the maximum EPS production (39.6gL(-1)) was appeared with maltose 48.65gL(-1), Mn(2+) 1118μM and Zn(2+) 901μM. The EPS also showed good H(2)O(2) scavenging activity while moderate free radical scavenging activity and reductive ability were determined. The EPS from WR-1 may be a new source of natural antioxidants with potential value for health, food and industry.     

Structural analysis of the extracellular polysaccharide produced by Sphaerotilus natans

An extracellular polysaccharide (EPS) was recovered and purified from the culture fluid of a sheathed bacterium, Sphaerotilus natans. Glucose, rhamnose, and aldobiouronic acid were detected in the acid hydrolysate of EPS by thin-layer chromatography (TLC). The aldobiouronic acid was found to be composed of glucuronic acid and rhamnose by TLC and gas-liquid chromatography analyses of the corresponding neutral disaccharide. The structure of EPS was identified by methylation linkage analysis and nuclear magnetic resonance. Additionally, partial acid hydrolysates of EPS were prepared and put through fast atom bombardment-mass spectrometry to determine the sugar sequence of EPS. The resulting data showed that EPS produced by S. natans is a new gellan-like polysaccharide constructed from a tetrasaccharide repeating unit, as shown below. -->4)-alpha-D-Glcp-(1-->2)-beta-D-GlcA p-(1-->2)-alpha-L-Rha p-(1-->3)-beta-L-Rha p-(1-->.     

Purification,structural characterization and antioxidant property of an extracellular polysaccharide from Aspergillus terreus

The marine fungus Aspergillus terreus produces an extracellular polysaccharide, YSS, when grown in potato dextrose-agar medium. YSS was isolated from the fermented liquids using ethanol precipitation, anion-exchange and size-exclusion chromatography. YSS was mainly composed of mannose and galactose in a molar ratio of 7.68:1.00, its average molecular weight was estimated to be about 18.6 kDa. On the basis of chemical and spectroscopic analyses, including one- and two-dimensional nuclear magnetic resonance (1D and 2D NMR) spectroscopy, structure of YSS may be represented, at an average, as a backbone of mannan with two types of branches. The mannan backbone is mainly composed of (1→2)-linked α-mannopyranose with small amounts of (1→6)-linked α-mannopyranose residues. The branches consist of terminal β-galactofuranose residues, and disaccharide units of (1→6)-linked α-mannopyranose. The branches are linked to C-6 of (1→2)-linked α-mannopyranose residues of backbone. The antioxidant activity of YSS was evaluated with the scavenging abilities on 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide and hydroxyl radicals in vitro, and the results indicated that YSS had good antioxidant activity, especially scavenging ability on DPPH radicals. The investigation demonstrated that YSS is a novel branched galactomannan with antioxidant activity, and differs from previously described extracellular polysaccharides.     

Isolation and characterization of exocellular polysaccharide produced byLactobacillus bulgaricus

Summary Lactobacillus bulgaricus strains grown on skim milk produce a viscosifying exocellular watersoluble heteropolysaccharide composed of galactose, glucose and rhamnose in an approximately molar ratio of 4:1:1. The molecular weight is approximately 500.000.     

Isolation and characterization of an extracellular polysaccharide from Pseudomonas caryophylli CFR 1705

Extracellular polysaccharides were isolated from Pseudomonas caryophylli CFR 1705 grown on lactose containing medium. The major fraction (no.1) obtained on DEAE-cellulose chromatography was composed of rhamnose, mannose and glucose in the ratio 1:3.26:4.97, respectively, and having a molecular weight of 1.1×10⁶ Da. Methylation followed by GC-MS analysis revealed it to be a highly branched 1,4-linked hexosan with mannose and glucose as the branch-off residues at positions C-2 and C-6 of the main chain. Rhamnose was essentially found as non-reducing terminal residue.     

Properties of an extracellular polysaccharide produced by a strain of enterobacter isolated from pond water

A bacterium which was isolated from pond water and identified as Enterobacter cloacae produced a viscous extracellular polysaccharide when it was grown aerobically in a medium containing sucrose as a sole source of carbon. The maximum molecular weight of the polysaccharide was about 9.0 x 10(5). The polysaccharide was composed of fucose, galactose, glucose, and glucuronic acid in a molar ratio of 2:3:2:1, but the molecular weight and the molar ratio of the sugar component were different from those of the polysaccharide produced by the same species reported elsewhere.     

Characterization of an extracellular polysaccharide produced by a Saharan bacterium Paenibacillus tarimensis REG 0201M

The purpose of this paper is to highlight the novel molecular characteristics and rheological properties of the polysaccharide produced by a bacterium isolated from extremophilic environment (Algerian Sahara). Phenotypic and molecular characteristics of the REG 0201M bacterial strain retained for this research were studied. Extracellular polysaccharide produced by REG 0201M was purified, characterized and its production was investigated. Analysis of 16S rDNA gene sequence showed that strain REG 0201M belonged to the species Paenibacillus tarimensis. In sucrose agar medium, 1.31 g dry weight of the polysaccharide per liter was produced. Evaluation of water absorption capacity showed that this polysaccharide was able to absorb 1000 times more water than its own weight. The average molecular weight determined by high-performance size exclusion chromatography multiangle laser light scattering was 1.718 × 106 g mol−1. Analysis of the monosaccharide composition by high-performance anion exchange chromatography showed the presence of fructose (77.67%) as the main neutral sugar, followed by galactose (20.37%), arabinose (1.79%), and rhamnose (0.16%). Its global charge determined by the Zeta potential measurement was about −35.27 ± 0.66 mV. The main functional groups were elucidated by Fourier-Transform Infrared Spectroscopy while the surface morphology was resolved by scanning electron microscopy analysis. Moreover, the rheological data revealed shear thinning properties with the same general behavior of the studied polysaccharide compared to xanthan. These results show the great potential of this polysaccharide, which could help to promote its use in various industries by replacing synthetic polymers.     

Chemical and rheological properties of an extracellular polysaccharide produced by the cyanobacterium Anabaena sp. ATCC 33047

The cyanobacterium (blue-green alga) Anabaena sp. ATCC 33047 produces an exopolysaccharide (EPS) during the stationary growth phase in batch culture. Chemical analysis of EPS revealed a heteropolysaccharidic nature, with xylose, glucose, galactose, and mannose the main neutral sugars found. The infrared (IR) spectrum of EPS showed absorption bands of carboxylate groups. The average molecular mass of the polymer was 1.35 MDa. Aqueous dispersions at EPS concentrations ranging from 0.2% to 0.6% (w/w) showed marked shear-thinning properties (power-law behavior). Linear dynamic viscoelastic properties showed that the elastic component was always higher than the viscous component. Viscous and viscoelastic properties demonstrated the absence of conformational changes within the concentration range studied. Stress-growth experiments revealed that 0.4% and 0.6% (w/w) EPS dispersions showed thixotropic properties. A detailed comparison of the linear dynamic viscoelasticity, transient flow, and decreasing shear rate flow curve properties was made for 0.4% (w/w) dispersions of xanthan gum (XG), Alkemir 110 (AG), and EPS. Viscoelastic spectra demonstrated that the EPS dispersion turned out to be more "fluidlike" than the AG and XG dispersions. The flow indexes indicated that the EPS dispersion was less shear-sensitive than that of XG, showing essentially the same viscosity, that is, >50 s(-1). The fact that viscosities of EPS and AG dispersions were not substantially different within the shear-rate range covered must be emphasized, in relation to EPS potential applications. The rheological behavior of EPS dispersions indicates the formation of an intermediate structure between a random-coil polysaccharide and a weak gel.