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

Juan Zhang; Yanqiu Cai; Guocheng Du; Jian Chen; Miao Wang; Zhen Kang

doi:10.1007/s12275-017-6514-4

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Evaluation and application of constitutive promoters for cutinase production by Saccharomyces cerevisiae: Juan Zhang ^1,2, Yanqiu Cai ¹, Guocheng Du ^2,3, Jian Chen ^2,4, Miao Wang ⁵, Zhen Kang ^1,2; Journal of Microbiology 2017;55(7):538-544
DOI: https://doi.org/10.1007/s12275-017-6514-4
Published online: June 30, 2017

¹The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, P. R. China, ²School of Biotechnology, Jiangnan University, Wuxi 214122, P. R. China, ³The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, P. R. China, ⁴National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, P. R. China, ⁵State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Jiangsu 214122, P. R. China¹The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, P. R. China, ²School of Biotechnology, Jiangnan University, Wuxi 214122, P. R. China, ³The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, P. R. China, ⁴National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, P. R. China, ⁵State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Jiangsu 214122, P. R. China

Corresponding author: Juan Zhang , Tel: +86-510-85918307,
Zhen Kang , Tel: +86-510-85918307,

Received: 10 December 2016 • Revised: 1 March 2017 • Accepted: 10 March 2017

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Abstract

died 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.

Keywords: cutinase; Thermobifida fusca; Saccharomyces cerevisiae; GAL1 system; constitutive promoter; TEF1

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

J. Microbiol. 2017;55(7):538-544. Published online June 30, 2017

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

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