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| A lipase-producing bacterium strain B68 was screened and identified as Pseudomonas fluorescens sp. Novel lipase gene lipB68 was cloned and sequenced… by the method of GenomeWalker. Sequence analysis revealed an open reading frame of 1,431 nucleotides, encoding a protein with 476 amino acids and a molecular mass of 50,193 Da, being a member of lipase subfamily I.3. The lipB68 gene was expressed in E. coli BL21 (DE3) in the form of inclusion body and the purification and protein refolding were conducted. Lipase lipB68 had a favorable reaction temperature of 20°C with p-Nitrophenyl caprate as substrate, lower than all the known lipases from subfamily I.3. And it also showed good stability to organic solvents such as DMF and acetonitrile. After being immobilized on fabric of cellulose, lipase lipB68 showed high enantioselectivity at the resolution of racemic a-phenylethanol and a-phenylpropanol (E=220, and 190 respectively). |
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| Keywords:clone, enzyme property, expression, lipase, Pseudomonas fluorescens |
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