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Gene Cloning and Characterization of a Novel Highly Organic Solvent Tolerant Lipase from Proteus sp. SW1 and its Application for Biodiesel Production |
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| Authors: | Wirongrong Whangsuk Pareenart Sungkeeree Sirinthra Thiengmag Jarunee Kerdwong Ratiboot Sallabhan Skorn Mongkolsuk Suvit Loprasert |
| Affiliation: | 1. Laboratory of Biotechnology, Chulabhorn Research Institute, Kamphaengphet 6 Road, Bangkok, 10210, Thailand 2. Applied Biological Sciences Program, Chulabhorn Graduate Institute, Bangkok, 10210, Thailand 3. Center of Excellence on Environmental Health and Toxicology, CHE, Ministry of Education, Bangkok, Thailand |
| Abstract: | Proteus sp. SW1 was found to produce an extracellular solvent tolerant lipase. The gene, lipA, encoding a bacterial lipase, was cloned from total Proteus sp. SW1 DNA. lipA was predicted to encode a 287 amino acid protein of 31.2?kDa belonging to the Group I proteobacterial lipases. Purified His-tagged LipA exhibited optimal activity at pH 10.0 and 55??C. It was highly stable in organic solvents retaining 112% of its activity in 100% isopropanol after 24?h, and exhibited more than 200% of its initial activity upon exposure to 60% acetone, ethanol, and hexane for 18?h. Biodiesel synthesis reactions, using a single step addition of 13% an acyl acceptor ethanol, showed that LipA was highly effective at converting palm oil into biodiesel. |
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