Isolation, Characterization and optimization of lipase produced from Bacteria

Amritha G Kulkarni, Ankalabassappa B Vedamurthy

Abstract


Lipasesare triacylglycerol acylhydrolases that catalyze the hydrolysis and thesynthesis of esters formed from glycerol and long chain fatty acids. Lipasesoccur widely in nature, but only microbial lipases are commerciallysignificant. Hence, isolation, characterization and optimization of lipaseenzyme was made by isolating lipase producing bacteria from oil contaminatedsoil around oil refinery industries, dairy industries, kitchen waste. Themorphological, cultural and biochemical characterization confirmed theorganisms as Bacillus spp, Staphylococcusaureus, Proteus mirabilis, Enterobacter freindii, Neisseria sp. andmolecular characterization of highest lipase producing bacteria was found to beBacillus subtilis KF015516.1 and wassubjected to further analysis .Bacillus subtilis wasoptimized using different parameters such as incubation period, carbon source,nitrogen source, temperature and pH. The activity of lipase enzyme of Bacillus subtilis showed highestactivity at 370c, pH of 8, NH4Cl as nitrogen source,carbon source of olive oil and castor oil among all the different conditionsprovided. The molecular weight of the lipase enzyme from Bacillus subtilisKF015516.1 was determined using SDS PAGE and it is found to be 45 kDa.Thedetergent activity of lipase enzyme from Bacillussubtilis KF015516.1 was analyzed and it was found that the lipase enzymealone does not have significant detergent property, however, when used alongwith the chemical detergent it showed the detergent activity. Hence, it can berecommended to be used as an additive in detergent preparation Key Words:  Lipase, Bacillussubtilis, detergent activity.

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ISSN : 2251-1563