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Cloning, expression and characterization of a new lipase from Yarrowia lipolytica

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Abstract

Bioinformatic analysis of the Yarrowia lipolytica CLIB122 genome has revealed 18 putative lipase genes all of which were expressed in Escherichia coli and screened for hydrolyzing activities against p-nitrophenyl-palmitate. One positive transformant containing an ORF of 1,098 bp encoding a protein of 365 amino acids was obtained. To characterize its enzymatic properties, the lipase gene was functionally expressed in Pichia pastoris. The resulting lipase exhibited the highest activity towards p-NP-decanoate at pH 7 and 35°C. In addition, the new lipase had a lower optimal temperature and pH compared to other Y. lipolytica lipases. It was noticeably enhanced by Ca2+, but was inhibited by PMSF, Hg2+ and Ni2+. The new lipase displayed the 1,3-specificity for triolein.

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Acknowledgments

We wish to thank Professor Ma, Institute of Microbiology, the Chinese Academy of Sciences, for donating the Y. lipolytica CGMCC 2.1405 strain. This work was supported by the National High Technology Research and Development Program of China (863 Program) (Nos. 2009AA03Z232, 2010AA101501), the Program for New Century Excellent Talents in University (NCET-07-0336), and the Natural Science Foundation of Hubei Province (2009CDA046).

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Authors and Affiliations

  1. Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

    Heyun Zhao, Xiaofeng Wang, Yun Liu, Li Xu & Yunjun Yan

  2. Laboratory of Structural Biology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

    Lina Zheng

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  1. Heyun Zhao
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  2. Lina Zheng
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  3. Xiaofeng Wang
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  4. Yun Liu
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  5. Li Xu
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  6. Yunjun Yan
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Correspondence to Yunjun Yan.

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Zhao, H., Zheng, L., Wang, X. et al. Cloning, expression and characterization of a new lipase from Yarrowia lipolytica . Biotechnol Lett 33, 2445–2452 (2011). https://doi.org/10.1007/s10529-011-0711-8

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  • DOI: https://doi.org/10.1007/s10529-011-0711-8

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