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Characterization of two novel lipase genes isolated directly from environmental sample

  • Biotechnologically Relevant Enzymes and Proteins
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Abstract

Two novel lipase genes (lipJ02, lipJ03) were isolated directly from environmental DNA via genome-walking method. Lipase gene lipJ02 contained an open reading frame (ORF) of 1,425 bp and encoded a 474-amino acids lipase protein, while lipase gene lipJ03 contained an ORF of 1,413 bp and encoded a 470-amino acids lipase protein. The lipase genes were cloned into expression vector pPIC9K and successfully integrated into a heterologous fungal host, Pichia pastoris KM71, and the recombinant P. pastoris were screened via a high-throughput method. The recombinants were induced by methanol to secrete active lipases into cultural medium. The recombinant lipases were also purified and characterized. The optimum temperature for the purified lipase LipJ02 and LipJ03 was 30 and 35°C, respectively, at pH 8.0. They exhibited similar thermostability, but LipJ02 exhibited better pH stability than LipJ03.

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Acknowledgements

The work was supported by the grant from the Ministry of Science and Technology, P.R. China (project No. 2002CCA400) and the grant from the Ministry of Education, P.R. China (project No. 20020251004).

We wish to thank Prof. Lixing Ma (Hubei University, P. R. China) for excellent technical assistance.

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, People's Republic of China

    Zhengbing Jiang, Hongping Wang, Yushu Ma & Dongzhi Wei

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  1. Zhengbing Jiang
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  2. Hongping Wang
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  3. Yushu Ma
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  4. Dongzhi Wei
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Correspondence to Yushu Ma or Dongzhi Wei.

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Jiang, Z., Wang, H., Ma, Y. et al. Characterization of two novel lipase genes isolated directly from environmental sample. Appl Microbiol Biotechnol 70, 327–332 (2006). https://doi.org/10.1007/s00253-005-0065-z

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  • DOI: https://doi.org/10.1007/s00253-005-0065-z

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