Summary: The gene, encoding a collagenase of was expressed in and purified by affinity chromatography. The transformant BL21 (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 of 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 of PrtV was 8·52. 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 °C and pH 7·5, respectively. The activity of PrtV was inhibited by chelators such as EDTA, EGTA and 1,10-phenanthroline; however, its activity was restored by the addition of various metal ions (Co, Mn, Ca, Cu, Niand Zn), indicating that PrtV is a metalloprotease. PrtV degraded both type I collagen and synthetic substrate FALGPA well, showing that PrtV is indeed a collagenase.


Article metrics loading...

Loading full text...

Full text loading...


Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error