Journal
JOURNAL OF BIOTECHNOLOGY
Volume 103, Issue 1, Pages 1-10Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0168-1656(03)00071-3
Keywords
catA; ccdA; Brevibacillus choshinensis; thioredoxin; thiol-disulfide oxidoreductase; epidermal growth factor
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Brevibacillus choshinensis (Bacillus brevis) is a protein-hyperproducing bacterium with a useful host-vector system for the production of recombinant proteins. Here, we cloned the ccdA-catA (CcdA associated thioredoxin-like thioldisulfide oxidoreductase) locus of B. choshinensis HPD31-S5. CatA protein (molecular weight, 19664) contains a thioredoxin-like motif, Cys-Gly-Pro-Cys. It was successfully expressed in B. choshinensis extracellularly (similar to 100 mug ml(-1) culture) using the secretion vector pNCMO2, and in Escherichia coli intracellularly (similar to 350 mug ml(-1) culture) with an amino-terminal His-tag. Both recombinant proteins showed thiol-disulfide oxidoreductase activity. Incubation of non-native human epidermal growth factor (hEGF) containing incorrect disulfide bonds with B. choshinensis cells secreting CatA protein resulted in the stimulation of the conversion of non-native hEGF to the native form. Furthermore, co-expression of CatA protein with recombinant hEGF in the B. choshinensis production system increased the yield of native hEGF. (C) 2003 Elsevier Science B.V. All rights reserved.
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