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Extracellular accumulation of recombinant protein by Escherichia coli in a defined medium

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Abstract

Extracellular accumulation of recombinant proteins in the culture medium of Escherichia coli is desirable but difficult to obtain. The inner or cytoplasmic membrane and the outer membrane of E. coli are two barriers for releasing recombinant proteins expressed in the cytoplasm into the culture medium. Even if recombinant proteins have been exported into the periplasm, a space between the outer membrane and the inner membrane, the outer membrane remains the last barrier for their extracellular release. However, when E. coli was cultured in a particular defined medium, recombinant proteins exported into the periplasm could diffuse into the culture medium automatically. If a nonionic detergent, Triton X-100, was added in the medium, recombinant proteins expressed in the cytoplasm could also be released into the culture medium. It was then that extracellular accumulation of recombinant proteins could be obtained by exporting them into the periplasm or releasing them from the cytoplasm with Triton X-100 addition. The tactics described herein provided simple and valuable methods for achieving extracellular production of recombinant proteins in E. coli.

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Acknowledgements

This research work was supported by China Postdoctoral Science Foundation (grant no. 20060400682) and by the Knowledge Innovation Program of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (grant no. 2007KIP405). I would like to thank my co-tutor Prof. Wei-Hong Jiang and Prof. Sheng Yang for supporting this research.

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  1. Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China

    Xiang-Yang Fu

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  1. Xiang-Yang Fu
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Fu, XY. Extracellular accumulation of recombinant protein by Escherichia coli in a defined medium. Appl Microbiol Biotechnol 88, 75–86 (2010). https://doi.org/10.1007/s00253-010-2718-9

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