Applied Microbiology and Biotechnology

, Volume 88, Issue 1, pp 75–86

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

Biotechnological Products and Process Engineering

DOI: 10.1007/s00253-010-2718-9

Cite this article as:
Fu, XY. Appl Microbiol Biotechnol (2010) 88: 75. doi:10.1007/s00253-010-2718-9

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.

Keywords

Chemical permeabilizationExtracellular productionEscherichia coliRecombinant proteinMembrane permeabilitySecretion

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghaiChina