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.
Keywords: disulfide formation; oxidative protein folding; redox potential; superoxidizing cells.
© 2014 Wiley Periodicals, Inc.