Separation of four extracellular proteases of<i>Vibrio parahaemolyticus</i>by polyacrylamide gel electrophoresis

The prtV gene, encoding a collagenase of Vibrio parahaemolyticus, was expressed in Escherichia coli and purified by affinity chromatography. The transformant E. coli BL2l (DE3)(pPRT2) secreted the recombinant Prtv, and the highest enzyme activity was detected in the culture supernatant after 5 h IPTG induction. The molecular mass o f purified PrtV was 62 kDa as determined by gel filtration, which was similar to that obtained by SDS-PAGE (64 kDa). This suggested that PrtV was a monomer protein having no subunit structure. The isoelectric point o f PrtV was 852. In addition, PrtV contained a 27 amino acid signal peptide, and the amino acid composition of the PrtV showed satisfactory agreement with that predicted from the DNA sequence. The optimum temperature and pH of PrtV were 40 OC and pH 7.5, respectively. The activity of PrtV was inhibited by chelators such as EDTA, EGTA and 1,lOphenanthroline; however, its activity was restored by the addition of various metal ions (Co2+, Mn2+, Ca2+, Cu2+, Ni2+ and Zn2+), indicating that PrtV is a metalloprotease. PrtV degraded both type I collagen and synthetic substrate FALGPA well, showing that PrtV is indeed a collagenase.

show abstract

scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.

Contact Info

customersupport@researchsolutions.com

10624 S. Eastern Ave., Ste. A-614

Henderson, NV 89052, USA

This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.

Blog Terms and Conditions API Terms Privacy Policy Contact Cookie Preferences Do Not Sell or Share My Personal Information

Made with 💙 for researchers

Part of the Research Solutions Family.

Separation of four extracellular proteases ofVibrio parahaemolyticusby polyacrylamide gel electrophoresis

Cited by 5 publications

References 9 publications

Cloning and sequence analysis of a novel metalloprotease gene from Vibrio parahaemolyticus 04

Cloning and sequence analysis of a novel metalloprotease gene from Vibrio parahaemolyticus 04

Evidence for the requirement of extracellular protease in the pathogenic interaction of Pyrenopeziza brassicae with oilseed rape

Expression and characterization of the prtV gene encoding a collagenase from Vibrio parahaemolyticus in Escherichia coli

Contact Info

Product

Resources

About