Azyme Biosciences Private Limited, Bangalore – 560069
Microbial lipases are currently receiving much attention with the rapid development of enzyme technology. Stability of Lipase enzymes with good activity has been an interesting field for researchers. Lipases are a class of enzymes which catalyze the hydrolysis of long-chain triglycerides. Production of extracellular lipase in submerged culture of Bacillus cereus MTCC 1272 strain has been investigated. The lipase production was optimized in shake flask experiments. Lipase enzyme was purified and molecular mass determined by SDS-PAGE. The purification steps included ammonium salt precipitation, dialysis and ion exchange chromatography on DEAE-cellulose. Enzyme exhibited high activity in the presence of calcium ion. Enzyme activity reduced in the presence of EDTA confirming enzyme to be a metalloprotein. The lipase enzyme found to be stable with high activity in boric acid and sodium taurocholate. Enzyme activity found to be increased more than 200% in salts and 150% in organic solvents like acetone and butanol. Enzyme was stable for more than 4 hours at 60°C to 75°C in salts and organic solvents. And the enzyme was also stable for more than 16 hours at room temperature. Combination of salts like boric acid and bile salts had shown high activity and stability at 80°C.
Key Words- Lipase, lipase activity, Salts and Organic Solvents, SDS PAGE, DEAE-cellulose