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Title

SCREENING, PURIFICATION AND PARTIAL CHARACTERIZATION OF A LIPASE FROM SOLVENT-TOLERANT BACTERIUM, STENOTROPHOMONAS SP. STRAIN IE-93

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 Start Page 169 | End Page 169

Abstract

 Organic solvent-tolerant bacteria are a novel and unique group of extremophilic microorganisms that can thrive in many harsh conditions. These bacteria are being explored for their potential in industrial applications, since their biocatalysis contents might be active in the presence of very high concentrations of organic solvents. In the present work, a screening programme was designed to isolate a novel methanol-tolerant bacterium. Among several different isolated bacteria, strain IE-93 was chosen as a best producer of lipase in the presence of 5% methanol. Phylogenetic characterization using 16S rDNA nucleotide sequence confirmed that the strain can be placed in STENOTROPHOMONAS genus and tentatively named STENOTROPHOMONAS sp. strain IE-93. The secreted lipase was purified by combination of amicon concentration, Q-Sepharose and Sephacryl S-200 column chromatography. The purified enzyme showed homogeneity on SDS-PAGE with apparent molecular mass of 60 kDa. The optimum pH and temperature for the enzyme were 8.0 and 35 oC respectively. The hydrolytic activity on various substrates (C4, C8, and C18 acyl chain) revealed that the purified enzyme has the highest priority for long acyl chains. Survey on lipolytic activity of the enzyme in the presence of various concentration of methanol (from 10% to 50%) showed its reasonable stability and more than 77% of initial activity was retained after 1 h incubation in 40% methanol. In overall, long alkyl chain substrate specificity and convenience stability in the presence of methanol suggest that the isolated lipase has biotechnological applications especially in low water content reactions and BIODIESEL production.

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