Abstract
Vibrio vulnificus is a ubiquitous estuarine microorganism but causes fatal systemic infections in immunocompromised humans, cultured eels or shrimps. An extracellular metalloprotease VVP/VvpE has been reported to be a potential virulence factor of the bacterium; however, a few strains isolated from a diseased eel or shrimp were recently found to produce a serine protease termed VvsA, but not VVP/VvpE. In the present study, we found that these strains had lost the 80 kb genomic region including the gene encoding VVP/VvpE. We also purified VvsA from the culture supernatant through ammonium sulfate fractionation, gel filtration and ion-exchange column chromatography, and the enzyme was demonstrated to be a chymotrypsin-like protease, as well as those from some vibrios. The gene vvsA was shown to constitute an operon with a downstream gene vvsB, and several Vibrio species were found to have orthologues of vvsAB. These findings indicate that the genes vvp/vvpE and vvsAB might be mobile genetic elements.
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Abbreviations
- Boc:
-
t-Butyloxycarbonyl
- CBB:
-
Coomassie brilliant blue R-250
- LB:
-
Luria–Bertani
- MCA:
-
4-Methylylcoumaryl-7-amide
- PAGE:
-
Polyacrylamide gel electrophoresis
- PCR:
-
Polymerase chain reaction
- PU:
-
Proteinase unit
- RT:
-
Reverse transcription
- Suc:
-
Succinyl
- VVH:
-
Vibrio vulnificus hemolysin
- VVP/VvpE:
-
Vibrio vulnificus protease
- VvsA:
-
Vibrio vulnificus serine protease
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Acknowledgments
This study was supported by a grant from the Program of Japan Initiative for Global Research Network on Infectious Diseases (J-GRID), the Ministry of Education, Culture, Sports, Science and Technology in Japan.
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Miyoshi, Si., Wang, J., Katoh, K. et al. An extracellular serine protease produced by Vibrio vulnificus NCIMB 2137, a metalloprotease-gene negative strain isolated from a diseased eel. World J Microbiol Biotechnol 28, 1633–1639 (2012). https://doi.org/10.1007/s11274-011-0969-y
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DOI: https://doi.org/10.1007/s11274-011-0969-y