Screening and Molecular Characterization of Extracellular Lipase Producing Bacillus Species from Coconut Oil Mill Soil

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Author(s) V.R. Gayathri | P. Perumal | Lini P Mathew | B. Prakash
Pages 502-509
Volume 2
Issue 7
Date July, 2013
Keywords Lipase, Bacillus spp., SDS PAGE and RAPD.

Lipases (triacylglycerol acylhydrolases E.C are ubiquitous enzymes of considerable physiological significance and industrial application. In view of this significance of lipase we performed screening and molecular characterization of extracellular lipase producing Bacillus species in soil samples collected from coconut oil mill. The dominant Lipase producing organisms were isolated from oil-spilled areas of the groundnut oil extracting industry at Salem. The isolates were identified as Bacillus spp, the influence of growth period on lipase production of Bacillus spp. (B1 – B5) was assessed by culturing it on media for 48hrs. Further, the influence of substrate concentration (2 to 4 %) on lipase production was also assessed on the optimized substrate, which maximized the lipase production. Isolated Bacillus spp.(B1 – B5) were assayed for extra cellular lipase production. One unit of lipase activity was defined as the amount of enzyme releasing one mole of free fatty acid in one minute under standard assay condition. The purification method consists of Ammonium Sulphate precipitation, Dialysis and Column chromatography and Molecular determination by SDS PAGE. In genotypic characterization, RAPD analysis was done to characterize the organism.

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