Journal of Microbiology

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Evaluation and application of constitutive promoters for cutinase production by Saccharomyces cerevisiae: Juan Zhang , Yanqiu Cai , Guocheng Du , Jian Chen , Miao Wang , Zhen Kang; 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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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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Functional characterization of the cutI gene for the transcription of carbon monoxide dehydrogenase genes in Mycobacterium sp. strain JC1 DSM 3803: Jae Ho Lee , Sae Woong Park , Young Min Kim , Jeong-Il Oh; J. Microbiol. 2017;55(1):31-36. Published online December 30, 2016; DOI: https://doi.org/10.1007/s12275-017-6572-7

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Abstract

Carbon monoxide dehydrogenase (CO-DH) in Mycobacterium sp. strain JC1 is a key enzyme for the carboxydotrophic growth, when carbon monoxide (CO) is supplied as a sole source of carbon and energy. This enzyme is also known to act as nitric oxide dehydrogenase (NO-DH) for the detoxification of NO. Several accessory genes such as cutD, cutE, cutF, cutG, cutH, and cutI, are clustered together with two copies of the CO-DH structural genes (cutB1C1A1 and cutB2C2A2) in Mycobacterium sp. strain JC1 and are well conserved in carboxydotrophic mycobacteria. Transcription of the CO-DH structural and accessory genes was demonstrated to be increased significantly by acidified sodium nitrate as a source of NO. A cutI deletion (ΔcutI) mutant of Mycobacterium sp. strain JC1 was generated to identity the function of CutI. Lithoautotrophic growth of the ΔcutI mutant was severely affected in mineral medium supplemented with CO, while the mutant grew normally with glucose. Western blotting, CO-DH activity staining, and CO-DH-specific enzyme assay revealed a significant decrease in the cellular level of CO-DH in the ΔcutI mutant. Northern blot analysis and promoter assay showed that expression of the cutB1 and cutB2 genes was significantly reduced at the transcriptional level in the ΔcutI mutant, compared to that of the wildtype strain. The ΔcutI mutant was much more susceptible to NO than was the wild type.

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Characterization of a MHYT domain-coupled transcriptional regulator that responds to carbon monoxide
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Nucleic Acids Research.2024; 52(15): 8849. CrossRef
Molybdenum Enzymes and How They Support Virulence in Pathogenic Bacteria
Qifeng Zhong, Bostjan Kobe, Ulrike Kappler
Frontiers in Microbiology.2020;[Epub] CrossRef

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