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Metal Content and Biochemical Analyses of a Recombinant Collagenase PrtV from Vibrio parahaemolyticus
Author(s) -
Yu Meishiuan,
Yap Meengan,
Lee Chiayin
Publication year - 2000
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2000.tb02567.x
Subject(s) - collagenase , vibrio parahaemolyticus , recombinant dna , substrate (aquarium) , vibrio alginolyticus , enzyme , biochemistry , hydrolysis , metal ions in aqueous solution , biology , extracellular , metalloproteinase , microbiology and biotechnology , vibrio , chemistry , metal , bacteria , ecology , genetics , organic chemistry , gene
PrtV is an extracellular metalloprotease of Vibrio parahaemolyticus and regarded as a collagenase. Inductively coupled plasma‐optical emission spectrometry analysis indicated that the recombinant PrtV contains 1 mol of zinc per mol of the native enzyme. On the basis of a kinetic study using 2‐furanacryloyl‐Leu‐Gly‐Pro‐Ala (FALGPA, the specific substrate for bacterial collagenase) as a substrate, it was suggested that metal ions may play a significant role in the binding and catalytic steps of the substrate. PrtV hydrolyzed type I, II, III, and IV collagens; however, it did not hydrolyze type V. In addition, the hydrolysis of native proteins and synthetic substrates revealed that PrtV possesses higher activity toward collagen and collagenlike sequences. The result of the thermal stability study indicated that PrtV was thermostable up to 40 C; at 50 C, stability gradually decreased. In addition, PrtV showed higher storage stability at −20 and 4 C, respectively, than at 25 C. Compared with collagenases from Clostridium histolyticum and Vibrio alginolyticus , PrtV was immunologically different and had no significant effect on the growth of CHO, HeLa, and Vero cells. Taken together, the results of the studies described in this paper advance our knowledge concerning the metal content and biochemical properties of PrtV.