Control of Folding of Proteins Secreted by a High Expression Secretion Vector, pIN-III-ompA: 16-Fold

Abstract

Denatured pro-subtilisin accumulated as aggregates in the Escherichia coli periplasmic space when pro-subtilisin was fused to the OmpA signal peptide using a high expression secretion vector, pIN-III-ompA, and only a small fraction of pro-subtilisin was properly folded to produce active subtilisin. Here we report a substantial improvement in the control of folding and a 16-fold increase of active subtilisin production by inducing the expression of the pre(ompA)-pro-subtilisin gene with a very low inducer concentration (0.005 mM IPTG) at a low culture temperature (23°C). These results indicate that the rate of protein synthesis and the culture temperature are important factors for proper folding of proteins secreted across the cytoplasmic membrane using pIN-III-ompA as vector.

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Takagi, H., Morinaga, Y., Tsuchiya, M. et al. Control of Folding of Proteins Secreted by a High Expression Secretion Vector, pIN-III-ompA: 16-Fold Increase in Production of Active Subtilisin E in Escherichia Coli. Nat Biotechnol 6, 948–950 (1988). https://doi.org/10.1038/nbt0888-948

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