PRODUCTION AND CHARACTERIZATION OF EXOPOLYSACCHARIDE FROM MARINE MODERATELY HALOPHILIC BACTERIUM HALOMONAS SMYRNENSIS SVD III
Keywords:Halomonas smyrnensis SVD III, Exopolysaccharide, Characterization of EPS by FTIR, MS, 1H NMR, Glycosidic linkages, Heptasaccharide, Moderately halophilic bacterium
Objective: To study 1) Optimization of nutritional and environmental parameters to enhance the yield of EPS by Halomonas smyrnensis SVD III isolated from seawater, West Coast of Maharashtra, India and 2) Purification and characterization of the EPS produced.
Methods: The isolate was grown in Sehgal and Gibbons (SG) medium broth supplemented with 3% glucose, at 37 Â°C, 120 rpm for 7 d. Optimization of different parameters was carried out with one factor at a time approach. EPS was isolated from cell-free supernatant of the culture broth by centrifugation and precipitation using chilled ethanol, after removal of proteins by trichloroacetic acid (TCA) treatment. Characterization of the purified EPS was carried out with respect to fourier-transform infrared (FTIR) spectrum, 1H nuclear magnetic resonance (NMR) spectrum and mass spectrometry (MS) analysis.
Results: Two-fold increase in the yield of EPS (23 g/l) by the selected isolate was obtained by using culture conditions as 10% inoculum size having cell density of 107 cells/ml, pH 6, incubation temperature 45 Â°C, 3% carbohydrate, 0.5% yeast extract as nitrogen source, 20% salt concentration and 7 d of incubation period. Characterization of the purified EPS suggested the presence of dominated glycosidic linkages and heptasaccharide nature of the molecule. As the present strain is halophilic, 20% NaCl was found to be optimum.
Conclusion: Optimization studies resulted in two-fold increase in the yield of EPS which is of heptasaccharide nature.
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