A. K. Paul

Production, Partial Purification and Some Bio-physicochemical Properties of EPS Produced by Halomonas xianhensis SUR308 Isolated from a Saltern Environment

Abstract A moderately halophilic bacterium, Halomonas SUR308, an efficient extracellular polymeric substance (EPS) producing strain was isolated from a multi-pond solar saltern of Odisha, India. The strain was identified as Halomonas xianhensis SUR308 (Genbank Accession No. KJ933394) based on 99 % similarity of 16S rRNA gene sequences of H. xianhensis. Kinetics of growth revealed that the isolate produced 2.56 g/L of EPS during mid-stationary phase when grown under continuous shaking (120 rpm). Chemical analysis of the partially purified SUR308-EPS has been shown to contain 97 % neutral sugar but no protein and uronic acid. Aqueous solution of EPS was highly viscous and displayed pseudoplastic and thixotropic behavior. It was also able to emulsify several hydrophobic substrates like oils and hydrocarbons efficiently. The scavenging effect of SUR308-EPS was correlated well with increasing concentrations and showed about 73 % DPPH radical scavenging at a concentration of 1 mg/ml. Effect of the EPS on human hepatocyte cells revealed no significant cytotoxic effect.

Production of Extracellular Polymeric Substances by Halophilic Bacteria of Solar Salterns

Moderately halophilic aerobic bacteria were isolated from 31 soil and 18 water samples collected from multipond solar salterns of Gujarat, Orissa, and West Bengal, India. A total of 587 bacterial isolates with distinct morphological features were obtained from these samples following dilution and plating on MH agar medium supplemented with NaCl. The isolates were screened for growth associated extracellular polymeric substances (EPS) production in MY medium under batch culture. In all, 20 isolates were selected as potent ones producing more than 1 g/L of EPS. These EPS producing isolates were characterized in detail for their morphological, physiological, and biochemical features and tentatively identified as members belonging to the genera Halomonas, Salinicoccus, Bacillus, Aidingimonas, Alteromonas, and Chromohalobacter. Apart from EPS production, these isolates also hold promise towards the production of various biomolecules of industrial importance.

Optimization of factors influencing exopolysaccharide production by Halomonas xianhensis SUR308 under batch culture

A moderately halophilic bacterium, Halomonas xianhensis SUR308 (GenBank Accession No. KJ933394) was isolated from multi-pond solar salterns of Odisha, India. Exopolysaccharide (EPS) production by this strain in malt extract yeast extract (MY) medium has been optimized under batch culture system. Among the different media tested, MY medium showed an EPS production of 2.55 g/L, which increased to 2.85 g/L under optimized aeration. An initial pH of 7.5 and incubation temperature of 32 °C were found to be most suitable for EPS production by the isolate under aerobic condition. An EPS production of 3.85 g/L was achieved when the growth medium was supplemented with 2.5% NaCl. Glucose was the most favourable carbon source for EPS production and maximum production (5.70 g/L) was recorded with 3% glucose. However, growth as well as production of EPS was remarkably affected when the growth medium was supplemented with hydrocarbons as sole source of carbon. Among different nitrogen sources, casein hydrolysate at 0.5% level was proved to be the best for EPS production and an initial inoculum dose of 7% (v/v) enhanced the EPS production to 7.78 g/L, while the divalent metal ions were in general toxic to growth and EPS production, EPS synthesis by SUR308 was enhanced with Cr (VI) supplementation.

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 (CIxrd, 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.