Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Purification and Properties of a Collagenolytic Protease Produced by Marine Bacterium Vibrio vulnificus CYK279H

  • Kang, Sung-Il (Department of Biotechnology & Bioengineering, Pukyong National University) ;
  • Jang, Young-Boo (Department of Biotechnology & Bioengineering, Pukyong National University) ;
  • Choi, Yeung-Joon (Division of Marine Bioscience/Institute of Marine Industry, Gyeongsang National University) ;
  • Kong, Jai-Yul (Department of Biotechnology & Bioengineering, Pukyong National University)
  • Published : 2005.12.31
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Abstract

A collagenolytic enzyme, produced by Vibrio vulnificus CYK279H, was purified by ultrafiltration, dialysis, Q-Sepharose ion exchange and Superdex-200 gel chromatography. The enzyme from the supernatant was purified 13.2 fold, with a yield of 11.4%. The molecular weight of the purified enzyme was estimated by SDS-PAGE to be approximately 35.0kDa. The N-terminal sequence of the enzyme was determined as Gly-Asp-Pro-Cys-Met-Pro-Ile-Ile-Ser-Asn. The optimum temperature and pH for the enzyme activity were $35^{\circ}C$ and 7.5, respectively. The enzyme activity was stable within the pH and temperature ranges 6.8-8.0 and $20{\sim}35^{\circ}C$, respectively. The purified enzyme was strongly activated by $Zn^{2+},\;Li^{2+},\;and\;Ca^{2+}$, but inhibited by $Cu^{2+}$. In addition, the enzyme was strongly inhibited by 1, 10-phenanthroline and EDTA. The purified enzyme was suggested to be a neutral metalloprotease.

Keywords

References

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