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