Purification and characterization of a protease produced by Vibrio mimicus.
A protease produced by Vibrio mimicus was purified to apparent homogeneity by ammonium sulfate fractionation and successive column chromatography on Sephacryl S-100 and Mono Q Monobeads. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) of the final preparation of the enzyme revealed the homogeneity of the purified enzyme. Conventional PAGE showed that the purified protease migrated as a single band with protease activity. The molecular weight of the protease was estimated to be about 31,000 on the basis of its mobility on sodium dodecyl sulfate-PAGE. The purified protease had both proteolytic and hemagglutination (HA) activities. The proteolytic and HA activities were inhibited by metalloprotease inhibitors and heat treatment. V. mimicus protease therefore appeared as a heat-labile, bifunctional molecule capable of mediating proteolysis and HA. The immunodiffusion analysis showed that the proteases produced by Vibrio cholerae and V. mimicus are immunologically cross-reactive.
Chowdhury
M A
MA
Department of Environmental Hygiene, Faculty of Pharmaceutical Sciences, Okayama University, Japan.
Miyoshi
S
S
Shinoda
S
S
D000596:Amino Acids / Q000032:analysis; D010450:Endopeptidases / Q000032:analysis / Q000302:isolation & purification* / Q000633:toxicity; D006388:Hemagglutinins / Q000032:analysis; D011480:Protease Inhibitors / Q000494:pharmacology; D014733:Vibrio / Q000201:enzymology* / Q000472:pathogenicity
D000596:Amino Acids; D006388:Hemagglutinins; D011480:Protease Inhibitors; D010450:Endopeptidases