| Title | Evaluation and application of constitutive promoters for cutinase production by Saccharomyces cerevisiae |
|---|---|
| Author | Juan Zhang1,2*, Yanqiu Cai1, Guocheng Du2,3, Jian Chen2,4, Miao Wang5, and Zhen Kang1,2* |
| Address | 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. China |
| Bibliography | Journal of Microbiology, 55(7),538–544, 2017, |
| DOI | 10.1007/s12275-017-6514-4 |
| Key Words | cutinase, Thermobifida fusca, Saccharomyces cerevisiae, GAL1 system, constitutive promoter, TEF1 |
| 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. |