Highly efficient folding of multi-disulfide proteins in superoxidizing Escherichia coli cytoplasm

Authors

  • Wenyao Zhang,

    1. Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, School of Pharmacy, East China University of Science and Technology, Shanghai, China
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  • Wenyun Zheng,

    1. Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, School of Pharmacy, East China University of Science and Technology, Shanghai, China
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  • Miaowei Mao,

    1. Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, School of Pharmacy, East China University of Science and Technology, Shanghai, China
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  • Yi Yang

    Corresponding author
    1. Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, School of Pharmacy, East China University of Science and Technology, Shanghai, China
    • Correspondence to: Y. Yang

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ABSTRACT

In this study, we monitored the thiol–disulfide redox potential of different Escherichia coli strains using redox-sensitive variants of green fluorescent protein. The cells with extreme oxidizing cytoplasm were generated by introducing a highly efficient disulfide relay system. The developed cells have exceptionally efficient de novo disulfide bond formation and significantly improve the oxidative folding of the client multi-disulfide proteins. Superoxidizing E. coli strain provides an effective method for the high-level production of recombinant disulfide-containing proteins. Biotechnol. Bioeng. 2014;111: 2520–2527. © 2014 Wiley Periodicals, Inc.

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