Immobilization of Organic Solvent-Tolerant Lipase from Pseudomonas mendocina M-37 with Potential Synthetic Activities

摘要

A thermostable solvent-tolerant lipase was isolated from Pseudomonas mendocina M-37. The lipase production medium was optimized for cost-effective production. Olive oil as a carbon source, and glycine as a nitrogen source were selected as the best for maximum lipase production. Medium optimization led to 3.75-fold increase in the lipase production. The extracellular lipase was purified 42.2-fold to homogeneity by precipitation using polyethyleneglycol, ultrafiltration and hydrophobic interaction chromatography. Its molecular mass, determined with sodium dodecyl sulphate-polyacrylamide gel electrophoresis, was 32 kDa. The enzyme was further immobilized on microcrystalline cellulose. The lipase showed an optimal water activity of 0.53 for both, acidolysis and interesterification reactions. Six- to sevenfold increase in synthetic activity of immobilized lipase was observed when interesterification activity of 0.139 IU/mg and transesterification activity of 0.181 IU/mg, respectively, were obtained. This is the first report on Pseudomonas mendocina lipase with synthetic activity immobilized on microcrystalline cellulose.