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Cloning, expression and characterization of a halotolerant esterase from a marine bacterium Pelagibacterium halotolerans B2T

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Abstract

An esterase PE10 (279 aa) from Pelagibacterium halotolerans B2T was cloned and overexpressed in Escherichia coli Rosetta in a soluble form. The deduced protein was 29.91 kDa and the phylogenetic analysis of the deduced amino acids sequence showed it represented a new family of lipolytic enzymes. The recombinant protein was purified by Ni–NTA affinity chromatography column and the characterization showed its optimal temperature and pH were 45 °C and pH 7.5, respectively. Substrate specificity study showed PE10 preferred short chain p-nitrophenyl esters and exhibited maximum activity toward p-nitrophenyl acetate. In addition, PE10 was a halotolerant esterase as it was still active under 4 M NaCl. Three-dimensional modeling of PE10 suggested that the high negative electrostatic potential on the surface may relevant to its tolerance to high salt environment. With this halotolerance property, PE10 could be a candidate for industrial use.

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Acknowledgments

This work was supported by the Open Research Program of the Key Laboratory of Marine Ecosystem and Biogeochemistry (LMEB) of the State Oceanic Administration (SOA) (grant LMEB201101).

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Authors and Affiliations

  1. College of Life Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou, 310058, Zhejiang, China

    Xiawei Jiang, Yingyi Huo, Hong Cheng, Xinqi Zhang, Xufen Zhu & Min Wu

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  1. Xiawei Jiang
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  2. Yingyi Huo
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  3. Hong Cheng
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  4. Xinqi Zhang
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  5. Xufen Zhu
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  6. Min Wu
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Correspondence to Min Wu.

Additional information

Communicated by A. Driessen.

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Jiang, X., Huo, Y., Cheng, H. et al. Cloning, expression and characterization of a halotolerant esterase from a marine bacterium Pelagibacterium halotolerans B2T . Extremophiles 16, 427–435 (2012). https://doi.org/10.1007/s00792-012-0442-3

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  • DOI: https://doi.org/10.1007/s00792-012-0442-3

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