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Identification and characterization of novel esterases from a deep-sea sediment metagenome

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Abstract

A deep-sea sediment metagenomic library was constructed and screened for lipolytic enzymes by activity-based approach. Nine novel lipolytic enzymes were identified, and the amino acid sequences shared 56% to 84% identity to other lipolytic enzymes in the database. Phylogenetic analysis showed that these enzymes belonged to family IV lipolytic enzymes. One of the lipolytic enzymes, Est6, was successfully cloned and expressed in Escherichia coli Rosetta in a soluble form. The recombinant protein was purified by Ni-nitrilotriacetic affinity chromatography column and characterized using p-nitrophenyl esters with various chain lengths. The est6 gene consisted of 909 bp that encoded 302 amino acid residues. Est6 was most similar to a lipolytic enzyme from uncultured bacterium (ACL67845, 61% identity) isolated from the South China Sea marine sediment metagenome. The characterization of Est6 revealed that it was a cold-active esterase and exhibited the highest activity toward p-nitrophenyl butyrate (C4) at 20°C and pH 7.5.

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Acknowledgments

This work was supported by grants from Public Science and Technology Research Funds Projects of Ocean (201005032-3), China Ocean Mineral Resources R & D Association (COMRA) Special Foundation (DYXM-115-01-3-01), and the National Natural Science Foundation of China (40806066).

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Authors and Affiliations

  1. College of Life Sciences, Zhejiang University, Room 209, 866 Yuhangtang Road, Hangzhou, 310058, China

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

  2. Laboratory of Marine Ecosystem and Biogeochemistry, State Oceanic Administration, Hangzhou, 310012, China

    Xuewei Xu & Yuehong Wu

  3. Second Institute of Oceanography, State Oceanic Administration, Hangzhou, 310012, China

    Xuewei Xu & Yuehong Wu

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  1. Xiawei Jiang
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  2. Xuewei Xu
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  3. Yingyi Huo
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  4. Yuehong Wu
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  5. Xufen Zhu
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  6. Xinqi Zhang
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  7. Min Wu
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Corresponding author

Correspondence to Min Wu.

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Communicated by Shung-Jiang Liu.

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Jiang, X., Xu, X., Huo, Y. et al. Identification and characterization of novel esterases from a deep-sea sediment metagenome. Arch Microbiol 194, 207–214 (2012). https://doi.org/10.1007/s00203-011-0745-2

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  • DOI: https://doi.org/10.1007/s00203-011-0745-2

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