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

被引:35
作者
Kim, SK
Yang, JY
Cha, J [1 ]
机构
[1] Pusan Natl Univ, Coll Nat Sci, Div Biol Sci, Pusan 609735, South Korea
[2] Pukyong Natl Univ, Coll Fishery Sci, Div Food Sci & Biotechnol, Pusan 608737, South Korea
关键词
bacterial metalloprotease; Vibrio parahaemolyticus; zinc-binding bite;
D O I
10.1016/S0378-1119(01)00882-4
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The metalloprotease gene (vppC) from Vibrio parahaemolyticus 04 has been cloned and sequenced. The vppC gene contains an open reading frame of 2442 nucleotides encoding a polypeptide of 8 14 amino acids with a calculated molecular mass of 89,833 Da. The predicted amino acid sequence of VppC containing a zinc metalloprotease HEXXH consensus motif displays extensive homology to the collagenase from Vibrio alginolyticus. The activity of the recombinant protease produced in Escherichia coli was examined by gelatin zymography and proteolytic activity assays. The substrate specificity study showed that the type I collagen and synthetic collagenase substrate carbobenzoxy-glcyl-L-prolyl-glycyl-L-prolyl-L-alanine were the best substrates, indicating that the cloned metalloprotease is indeed a collagenase. Multiple alignment analysis of the amino acid sequences and the enzymatic properties such as molecular mass and substrate specificity revealed three distinct classes of Vibrio metalloproteases. The identification of a new metalloprotease gene expands the role of Vibrio metalloproteases as a virulence factor for host infection. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
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页码:277 / 286
页数:10
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