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Fervidobacterium changbaicum Lip1: identification, cloning, and characterization of the thermophilic lipase as a new member of bacterial lipase family V

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Abstract

A novel lipase gene encoded 315 amino acid residues was obtained using lipase-prospecting primers and genome walking from hyperthermophilic bacterium Fervidobacterium changbaicum CBS-1. Sequence alignment and phylogenetic analysis revealed this novel lipase is a new member of bacterial lipase family V. The recombinant enzyme F. changbaicum lipase 1 (FCLip1) showed maximum activity at 78°C and pH 7.8. It displayed extreme thermostability at 70°C and was also stable across a wide pH range from 6.0 to 12.0. Kinetic study demonstrated FCLip1 preferentially hydrolyzed middle-length acyl chains, especially p-nitrophenyl caprate and tricaprylin. With p-nitrophenyl caprate as a substrate, the enzyme exhibited a K m and k cat of 4.67 μM and 22.7/s, respectively. In addition, FCLip1 was resistant to various detergents and organic solvents. This enzyme is the first reported thermophilic lipase from bacterial family Thermotogaceae. Its extreme stability with respect to temperature and pH, along with its triglyceride hydrolysis activity, indicate that FCLip1 has high potential for future application.

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Acknowledgments

This work was supported by grants from the High Technology Program of China (# 2010AA101501) and the Natural Science Foundation of China (# 30970632, # 30821005).

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Authors and Affiliations

  1. Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun, 130023, China

    Jingang Cai, Yuan Xie, Bo Song, Yanping Wang, Zuoming Zhang & Yan Feng

  2. Key Laboratory of MOE for Microbial Metabolism and School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China

    Yuan Xie & Yan Feng

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  1. Jingang Cai
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  2. Yuan Xie
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  3. Bo Song
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  4. Yanping Wang
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  5. Zuoming Zhang
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  6. Yan Feng
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Corresponding author

Correspondence to Yan Feng.

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Fig. S1

Ramachandran plot of the final model from FCLip1 calculated with PROCHECK. Residues in most favored regions located in [A, B, L]; residues in additional allowed regions located in [a, b, l, p]; residues in generously allowed regions located in [~a, ~b, ~l, ~p]; residues in disallowed regions are unshaded. Glycine residues shown as closed triangles (DOC 38 kb)

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Cai, J., Xie, Y., Song, B. et al. Fervidobacterium changbaicum Lip1: identification, cloning, and characterization of the thermophilic lipase as a new member of bacterial lipase family V. Appl Microbiol Biotechnol 89, 1463–1473 (2011). https://doi.org/10.1007/s00253-010-2971-y

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  • DOI: https://doi.org/10.1007/s00253-010-2971-y

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