ISSN: 2249–9504
CAS CODEN: IJPCDX

MUTATIONAL STUDY OF BACILLUS SPECIES FOR PRODUCTION,PURIFICATION AND CHARACTERISATION OF LIPASE

Abstract

Author(s): Ashajyothi Chavan, Deepali Chougale, RamachandraYarapa Lakshmikantha*and S. Padmalatha RaiSatwadi

Lipase being a hydrolyse enzyme acts on lipids to give fatty acids and glycerol. Soil microbes were isolated and screened for the potential producers of lipase using Tributyrine agar plate method. Bacillus lichiniformisMTCC2617 is optimised for comparative study with wild microorganism. Wild and MTCC2617 are inoculated in the production medium contained standardised carbon and nitrogensource. Wild was UV mutated for the strain improvement. The activity of the wild was increased from 780 U/ml to 2120 U/mlfor 20mintues incubation. The crude enzyme from wild, mutant wild and MTCC2617 was partially purified by ammonium sulphate precipitation, dialysis and ion exchange chromatography. After the complete purification percentage of yield was high for wild mutated as compare to MTCC2617. The molecular weight of the protein was found to be 55KDa for both MTCC2617 and mutant wild. Characterisation of the lipase enzymes showed optimum pH 9.0 for wild, mutant wild and 7.0 for MTCC2617. Optimum temperature was 550C. The enzyme was activated by 9μM of Calcium chloride and in the presence of 2.5μM Ethylenediamine tetra acetic acid the activity was reduced to 12.5% for mutated and MTCC 2617.

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