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Evaluation and application of constitutive promoters for cutinase production by Saccharomyces cerevisiae

  • Microbial Genetics, Genomics and Molecular Biology
  • Published:
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Abstract

Cutinase as a promising biocatalyst has been intensively studied and applied in processes targeted for industrial scale. In this work, the cutinase gene tfu from Thermobifida fusca was artificially synthesized according to codon usage bias of Saccharomyces cerevisiae and investigated in Saccharomyces cerevisiae. Using the α-factor signal peptide, the T. fusca cutinase was successfully overexpressed and secreted with the GAL1 expression system. To increase the cutinase level and overcome some of the drawbacks of induction, four different strong promoters (ADH1, HXT1, TEF1, and TDH3) were comparatively evaluated for cutinase production. By comparison, promoter TEF1 exhibited an outstanding property and significantly increased the expression level. By fed-batch fermentation with a constant feeding approach, the activity of cutinase was increased to 29.7 U/ml. The result will contribute to apply constitutive promoter TEF1 as a tool for targeted cutinase production in S. cerevisiae cell factory.

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Authors and Affiliations

  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, P. R. China

    Juan Zhang, Yanqiu Cai & Zhen Kang

  2. School of Biotechnology, Jiangnan University, Wuxi, 214122, P. R. China

    Juan Zhang, Guocheng Du, Jian Chen & Zhen Kang

  3. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, P. R. China

    Guocheng Du

  4. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, P. R. China

    Jian Chen

  5. State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Jiangsu, 214122, P. R. China

    Miao Wang

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  1. Juan Zhang
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  2. Yanqiu Cai
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  3. Guocheng Du
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  4. Jian Chen
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Correspondence to Juan Zhang or Zhen Kang.

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Zhang, J., Cai, Y., Du, G. et al. Evaluation and application of constitutive promoters for cutinase production by Saccharomyces cerevisiae . J Microbiol. 55, 538–544 (2017). https://doi.org/10.1007/s12275-017-6514-4

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  • DOI: https://doi.org/10.1007/s12275-017-6514-4

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