Cloning and characterization of the gene (empV) encoding extracellular metalloprotease from Vibrio vulnificus

Gene
Y C ChuangM C Chang

Abstract

A gene (empV) encoding the extracellular metalloprotease of Vibrio vulnificus CKM-1 has been cloned and sequenced. When the empV gene was expressed in minicells, a unique peptide of approx. 46 kDa was identified. Protease activity staining experiments also indicated a similar M(r) for the protease. The empV gene product (EmpV) is secreted into the periplasm of Escherichia coli, but not out of it. The crude enzyme prepared from the periplasmic fraction of recombinant E. coli was inhibited by a metalloprotease inhibitor and Zn2+ is essential for its protease activity. Nucleotide sequence analysis predicted a single open reading frame (ORF) of 1818 bp encoding a 606 amino acid (aa) polypeptide, with a potential 24 aa signal peptide followed by a long 'pro' sequence consisting of 172 aa. The N-terminal 20 aa sequence for the elastolytic protease (EepV), purified from the culture supernatant of V. vulnificus ATCC 29307, completely identified the beginning of the predicted mature protein within the deduced aa sequence except for 1 aa residue difference. The estimated pI and molecular weight of the predicted mature protein were 5.86 and 44.3 kDa, respectively, which are nearly identical to those of V. vulnificus L-180 extracellular ne...Continue Reading

References

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Citations

Aug 2, 2007·Journal of Industrial Microbiology & Biotechnology·Hyun-Soo KimKenneth N Timmis
Jun 28, 2008·The Protein Journal·Ilya V DemidyukSergey V Kostrov
Apr 1, 2000·Microbes and Infection·M S Strom, R N Paranjpye
Oct 18, 2002·Antimicrobial Agents and Chemotherapy·Hung-Jen TangYin-Ching Chuang
Oct 31, 2002·Applied and Environmental Microbiology·Katsushiro MiyamotoHiroshi Tsujibo
Nov 2, 2004·BMC Microbiology·Steven M DenkinDavid R Nelson
Oct 22, 2004·Emerging Infectious Diseases·Po-Ren HsuehYin-Ching Chuang
Sep 14, 2001·Biochemical and Biophysical Research Communications·J H SuM C Chang
May 10, 2002·Bioscience, Biotechnology, and Biochemistry·Katsushiro MiyamotoYoshihiko Inamori
Apr 5, 2005·Archaea : an International Microbiological Journal·Sebastian BäumerUwe Deppenmeier
May 17, 2011·International Journal of Medical Microbiology : IJMM·Yu-Chung Chen, Ya-Ting Chung
May 28, 2003·Extremophiles : Life Under Extreme Conditions·Cristina Sánchez-PorroAntonio Ventosa
Jul 31, 1999·Applied and Environmental Microbiology·S M Denkin, D R Nelson
Nov 5, 1997·Applied and Environmental Microbiology·T M ChangM C Chang

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