Ethanol effects on the overexpression of heterologous catalase in Escherichia coli BL21 (DE3)
- 34 Downloads
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.
KeywordsEthanol treatment Escherichia coli Heterologous protein expression Maltose ABC transporter prpD
We would like to thank Dr. Changhao Bi for CRISPR/cas9 method to knockout target genes from E. coli genome.
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).
Compliance with ethical standards
Conflict of interests
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Babu M, Bundalovic-Torma C, Calmettes C, Phanse S, Zhang Q, Jiang Y, Minic Z, Kim S, Mehla J, Gagarinova A, Rodionova I, Kumar A, Guo H, Kagan O, Pogoutse O, Aoki H, Deineko V, Caufield JH, Holtzapple E, Zhang Z, Vastermark A, Pandya Y, Lai CC, El Bakkouri M, Hooda Y, Shah M, Burnside D, Hooshyar M, Vlasblom J, Rajagopala SV, Golshani A, Wuchty S, J FG, Saier M, Uetz P, T FM, Parkinson J, Emili A (2018) Global landscape of cell envelope protein complexes in Escherichia coli. Nat Biotechnol 36(1):103–112. https://doi.org/10.1038/nbt.4024 CrossRefPubMedGoogle Scholar
- Goff J, Yee N (2017) Tellurate enters Escherichia coli K-12 cells via the SulT-type sulfate transporter CysPUWA. FEMS Microbiol Lett 364(24): fnx241. https://doi.org/10.1093/femsle/fnx241
- Lochowska A, Iwanicka-Nowicka R, Zaim J, Witkowska-Zimny M, Bolewska K, Hryniewicz MM (2004) Identification of activating region (AR) of Escherichia coli LysR-type transcription factor CysB and CysB contact site on RNA polymerase alpha subunit at the cysP promoter. Mol Microbiol 53(3):791–806. https://doi.org/10.1111/j.1365-2958.2004.04161.x CrossRefPubMedGoogle Scholar
- Sirko A, Zatyka M, Sadowy E, Hulanicka D (1995) Sulfate and thiosulfate transport in Escherichia-Coli K-12 - evidence for a functional overlapping of sulfate-binding and thiosulfate-binding proteins. J Bacteriol 177(14):4134–4136. https://doi.org/10.1128/jb.177.14.4134-4136.1995 CrossRefPubMedPubMedCentralGoogle Scholar
- Yu Z, Zheng H, Zhao X, Li S, Xu J, Song H (2016) High level extracellular production of a recombinant alkaline catalase in E. coli BL21 under ethanol stress and its application in hydrogen peroxide removal after cotton fabrics bleaching. Bioresour Technol 214:303–310. https://doi.org/10.1016/j.biortech.2016.04.110 CrossRefPubMedGoogle Scholar
- Zou C, Duan X, Wu J (2014) Enhanced extracellular production of recombinant Bacillus deramificans pullulanase in Escherichia coli through induction mode optimization and a glycine feeding strategy. Bioresour Technol 172:174–179. https://doi.org/10.1016/j.biortech.2014.09.035 CrossRefPubMedGoogle Scholar