Purification and characterization of the lipase enzyme from Bacillus megaterium M22

Lipases (triacylglycerol acylhydrolase, EC. 3.1.1.3), have a wide application field in industry depending on to act in micro-aqueous environment and the capabilities of catalyze esterification, transesterification (interesterification, aminolysis, acidolysis, alcoholysis) reactions. Lipases have gained special interest in recent years because of their features such as the facilitations production of special compounds which has chemically diffucult to synthesis with stereo and regional specific features, being specific to ester bond, the capabilities of using a wide spectrum of substrates, preventing the formation of side products, not to need cofactor to catalyze hydrolytic reactions and protections activity in organic solvents. In this study, lipase enzyme from a new Bacillus megaterium M22 isolated from soil was purified and made characterization studies. The lipase enzyme was purified 4 fold with % 34,42 yield using ammonium sulfate precipitation and DEAE-cellulose anion exchange chromatography. The molecular mass of lipase enzyme was identified 45 kDa with SDS-PAGE. The lipase was purified from Bacillus megaterium M22, were determined as the optimum temperature is 40 °C and the optimum pH value is 7,0. Methanol, toluene, benzene and chlorobenzene greatly enhanced the activity of lipase. Various metals were observed to reduce the activity of the lipase. The effect of various reagents on the lipase enzyme also was examined. Michaelis-Menten kinetic constants of the lipase enzyme which were purified from Bacillus megaterium M22, Km and Vmax values of 4.74 µM and 8.13 U/ml respectively were determined.