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Cloning, expression, and biochemical characterization of a thermostable lipase from Geobacillus stearothermophilus JC

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Abstract

A thermophilic lipase gene of Geobacillus stearothermophilus JC was cloned and expressed in a pET 28-a (+) expression vector. The biochemical properties of the recombinant enzyme and its enantioselective hydrolysis of (RS)-1-phenylethyl acetate were studied. Removal of the signal peptide greatly increased the enzyme’s expression level by 4.3 times. The purified JC lipase had an optimum temperature of 55°C and optimum pH of 9. Furthermore, comparisons with other enzymes suggest that a few amino acid alterations may significantly change the thermostability of this enzyme. The hydrolysis of (RS)-1-phenylethyl acetate with the crude recombinant JC lipase at 25°C produce (R)-1-phenylethanol in 97.7% e.e. and 46.1% yield after 24 h, corresponding to an E value of 237.

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Acknowledgments

This work was supported by a key project from Science & Technology Department of Zhejiang Province (2007C21150).

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The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Center for Biomedicine and Health, Hangzhou Normal University, 222 Wenyi Road, 310012, Hangzhou, China

    Yu Jiang & Zhenming Chen

  2. College of Biological & Environmental Engineering, Zhejiang University of Technology, 310032, Hangzhou, China

    Yu Jiang & Xiaoyun Zhou

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  1. Yu Jiang
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  2. Xiaoyun Zhou
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  3. Zhenming Chen
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Correspondence to Zhenming Chen.

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Jiang, Y., Zhou, X. & Chen, Z. Cloning, expression, and biochemical characterization of a thermostable lipase from Geobacillus stearothermophilus JC. World J Microbiol Biotechnol 26, 747–751 (2010). https://doi.org/10.1007/s11274-009-0213-1

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  • DOI: https://doi.org/10.1007/s11274-009-0213-1

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