Cloning, expression and characterisation of lipase from Anoxybacillus sp. Pdf1

Lipases (triacylglycerol acylhydrolases; EC.3.1.1.3) catalyze the hydrolysis of triglycerides at the oil-water interface. Microbial lipases have currently received considerable attention with regard to biotechnological applications for detergents, transesterification and chiral compound synthesis. In particular, lipases from thermophiles are expected to play a significant role in industrial processes, since they are thermostable and resistant to chemical denaturation.This work describes the cloning, isolation, and characterization of lipase from Anoxybacillus sp. PDF1. A 740 nt lipase gene coding for lipase from Anoxybacillus sp. PDF1 sequenced and cloned into pET28a+ vector. Vector encoding this gene was transformed into E.coli BL21, expressed and purified. The purified lipase showed optimal activity at 60°C in pH 8. It was determined that the enzyme had a lower Km (0,348 mM) for paranitrophenylbutyrate than most of the lipases. Metal ion effects to lipase were observed with cloride salts of Mg+2, Li, Ca+2, K, Zn+2, ve Co+metal ions. The molecular mass of the lipase was determined to be 24 kDa on SDS-PAGE. In the light of all data it has been suggested that the enzyme?s biocatalytic properties proved to be one of the important industrial enzymes.