Abstract
A novel method involving ethanol-induced increase in the heterologous recombinant protein expression in E. coli cells was commonly used in recent studies. However, the detailed mechanism of this method is still to be revealed. This work used comparative transcriptomic analysis and numerous experiments to uncover the mechanism of ethanol effects on the expression of heterologous catalase in the recombinant strain E. coli BL21 (pET26b-katA). The key regulatory genes malK and prpD were found to have the most significant effects on the expression of heterologous catalase. Thus, the maltose ABC transporter and carbon metabolism from propanoate metabolism to citrate cycle were found to be the main regulatory pathways activated by ethanol to enhance the synthesis of heterologous proteins. Based on these mechanisms, a universally applicable E. coli expression host strain for improving the expression of heterologous proteins might be constructed.
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Acknowledgments
We would like to thank Dr. Changhao Bi for CRISPR/cas9 method to knockout target genes from E. coli genome.
Funding
This study was supported by the foundation (Grant 2017KF005) of Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education and Tianjin Key Laboratory of Industrial Microbiology (Tianjin University of Science &Technology). And this work was also supported by the National Natural Science Fund of China (Grant 31701534), the Key Deployment Project in Chinese Academy of Sciences (Grant KFJ-STS-ZDTP-016-1, KFZD-SW-211-2), the Tianjin Science & Technology Planning Project (Grant 16YFZCSY00790, 15PTCYSY00020, and 15YFYSSY00040) and Yantai Marine economy innovation development demonstration project (Grant YHCX-SW-L-201703).
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Zheng, H., Yu, Z., Shu, W. et al. Ethanol effects on the overexpression of heterologous catalase in Escherichia coli BL21 (DE3). Appl Microbiol Biotechnol 103, 1441–1453 (2019). https://doi.org/10.1007/s00253-018-9509-0
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DOI: https://doi.org/10.1007/s00253-018-9509-0