title: Purification and characterization of Vibrio vulnificus protease.
abstract: A protease was purified from a strain of Vibrio vulnificus isolated from the blood of a septicemic human. The vibrio was cultured in bacto peptone-yeast extract medium, and the protease was purified by a purification procedure including ultrafiltration of the culture supernatant with an Amicon YM 5 membrane, diethylaminoethyl-Sephacel column chromatography, Sephacryl S-200 column chromatography and fast protein liquid chromatography on Mono Q column. The protease preparation revealed homogeneity on polyacrylamide gel electrophoresis and about 30,000-fold purification was achieved, with a yield of about 30%. The isoelectric point of the purified V. vulnificus protease was about 5.80 and its molecular weight was ca. 45,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH of the protease activity was 8.0. The V. vulnificus protease was inhibited by a metalloprotease inhibitor and zinc ion and/or ferrous ion were essential for its enzyme activity. No cysteine residue was detected in the V. vulnificus protease. The protease had caseinolytic, elastolytic and collagenolytic activities.
authors.forename: N
authors.initials: N
authors.lastname: Miyoshi
authors.forename: C
authors.initials: C
authors.lastname: Shimizu
authors.forename: S
authors.initials: S
authors.lastname: Miyoshi
authors.forename: S
authors.initials: S
authors.lastname: Shinoda
mesh_terms: D000596:Amino Acids / Q000032:analysis; D002614:Chelating Agents / Q000494:pharmacology; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D014733:Vibrio / Q000201:enzymology*
chemical_list: D000596:Amino Acids; D002614:Chelating Agents; D008670:Metals; D011480:Protease Inhibitors; D010447:Peptide Hydrolases