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Updated: Oct 2, 2026
Phage Phenomics: Physiological Approaches to Characterize Novel Viral Proteins
Published on: June 11, 2015
Cloning and sequence analysis of a novel metalloprotease gene from Vibrio parahaemolyticus 04
Soo Kwang Kim1, Ji Young Yang, Jaeho Cha
1Division of Biological Sciences, College of Natural Sciences, Pusan National University, Pusan 609-735, South Korea.
Abstract:
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 814 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-glycyl-L-prolyl-glycyl-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.
Insights
Researchers cloned and sequenced the Vibrio parahaemolyticus metalloprotease gene (vppC), identifying it as a collagenase. This discovery expands the understanding of Vibrio metalloproteases as potential virulence factors in host infections.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Vibrio species are known to produce metalloproteases.
- Metalloproteases can play significant roles in bacterial virulence and host tissue degradation.
Purpose of the Study:
- To clone and characterize the metalloprotease gene (vppC) from Vibrio parahaemolyticus.
- To determine the enzymatic activity and substrate specificity of the VppC protein.
- To understand the evolutionary relationships among Vibrio metalloproteases.
Main Methods:
- Gene cloning and sequencing of vppC from Vibrio parahaemolyticus.
- Recombinant protein expression in Escherichia coli.
- Gelatin zymography and proteolytic activity assays.
- Substrate specificity analysis using type I collagen and synthetic substrates.
- Multiple sequence alignment and phylogenetic analysis.
Main Results:
- The vppC gene encodes an 814-amino acid protein (VppC) with a molecular mass of 89,833 Da.
- VppC exhibits homology to known collagenases and possesses zinc metalloprotease activity.
- The enzyme efficiently degrades type I collagen and synthetic collagenase substrates.
- Analysis revealed three distinct classes of Vibrio metalloproteases.
Conclusions:
- The cloned metalloprotease from Vibrio parahaemolyticus is a collagenase.
- The VppC enzyme represents a novel virulence factor for Vibrio species.
- This finding contributes to the understanding of Vibrio pathogenesis and metalloprotease diversity.
