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

Praveen Dahiya, Subhash Chand2 and Neeraj Dilbaghi1*

Department of Bio- and Nanotechnology, Guru Jambheshwar University of Science and Technology, Hisar-125001, Haryana, India
2Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology, Hauz Khas, New Delhi-110016, India
§Present address: Amity Institute of Biotechnology, Amity University, Sector 125, Gautam Buddha Nagar, Noida-201303, Uttar Pradesh, India

Article history

Received May 27, 2013
Accepted March 17, 2014

Key words

acidolysis, immobilization, organic solvent-tolerant lipase, Pseudomonas mendocina, purification


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.

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