RT Journal Article SR Electronic(1) A1 DEANE, S. M. A1 MAHARAJ, R. A1 ROBB, F. T. A1 WOODS, D. R.YR 1987 T1 Cloning, Expression and Release of a Vibrio alginolyticus SDS-resistant Ca2+-dependent Exoprotease in Escherichia coli JF Microbiology, VO 133 IS 8 SP 2295 OP 2302 DO https://doi.org/10.1099/00221287-133-8-2295 PB Microbiology Society, SN 1465-2080, AB SUMMARY: A Vibrio alginolyticus SDS-resistant, Ca2+-dependent serine exoprotease gene, cloned on a 7-1 kb DNA fragment in the recombinant plasmid pVP100, was expressed from its own promoter in Escherichia coli. Although active exoprotease was produced by late stationary phase E. coli(pVP100) cultures after 15 h incubation in proteinaceous medium containing Ca2+, transcription and translation of the exoprotease occurred before 6 h, during exponential growth. The cloned exoprotease was synthesized as a pool of inactive precursor molecules during exponential growth, and released as active exoprotease 8 h later by a process which did not require protein synthesis or involve cell lysis. Release of the exoprotease by E. coli(pVP100) was not inhibited by o-phenanthroline, quinacrine or cerulenin. Supernatant samples from E. coli(pVP100) cultures contained two SDS-resistant exoproteases with apparent M r values of approximately 54000 and 39000. The cloned exoprotease activity was inhibited by EDTA and a serine protease inhibitor, but was not affected by an inhibitor of trypsin-like enzymes., UL https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-133-8-2295