Purification of a Serine Protease of Vibrio parahaemolyticus and Its Characterization

Abstract

A 50 kDa protease designated as VPP1 was purified from the culture supernatant of a clinical strain of Vibrio parahaemolyticus by ammonium sulfate fractionation, Sephacryl S-200 HR gel filtration and Fractogel EMD TMAE 650 ion-exchange chromatography. VPP1 was inhibited by EDTA, EGTA and serine protease inhibitors, suggesting that it is a calcium-dependent serine protease. N-terminal amino acid sequence of VPP1 was quite similar to that of V. metschnikovii protease and antibody against VPP1 inhibited the activity of V. metschnikovii protease, suggesting the similarity of the two proteases. It was demonstrated that VPP1 or its related protease widely distribute in not only V. parahaemolyticus but also V. alginolyticus.

Identification of Lys-Pro-Gln as a Novel Cleavage Site Specificity of Saliva-associated Proteases

“…The inhibitory effect of EDTA (65.3%) is pointing toward the importance of metal ions for activity. Indeed, metallo as well as some serine proteases have been reported to be metal ion-dependent (40,41). The hydrolysis of KPQ-pNA could be completely abolished by the combination of PMSF and EDTA (data not shown).…”

Photobacterium damselae subsp. damselae, an Emerging Pathogen Affecting New Cultured Marine Fish Species in Southern Spain

“…damselae strains and their virulence properties could not also be established . Several extracellular bacterial proteases, mainly metalloproteases and serine-proteases such as vibriolysins, are considered as virulence factors in numerous bacterial pathogens (Ishihara et al, 2002;Miyoshi et al, 2002;Farto et al, 2006). These proteases provoke tissue damages and degradation of host tissues, favouring the colonization and invasion of pathogens into the host (Miyoshi & Shinoda, 2000).…”

Generation of active fragments from human zymogens in the brady kinin-generating cascade by extracellular proteases from Vibrio vulnificus and V. parahaemolyticus

“…V. parahaemolyticus serine protease (50-kDa) was purified according to the procedures reported by Ishihara et al (2002).…”

“…However, the mechanism by which V. vulnificus metalloprotease can convert human zymogens is not clarified yet. Some other human pathogenic Vibrio species including V. parahaemolyticus, an etiologic agent causing watery diarrhea, secretes serine proteases (Ishihara et al, 2002;Lee et al, 2002). Although this serine protease can digest various host proteins, its biological or pathogenic actions are not documented.…”