Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of the Fibrinolytic Enzyme Produced by Bacillus subtilis KCK-7 from Chungkookjang

  • Paik, Hyun-Dong (Department of Food Science and Biotechnolgy of Animal Resources, Konkuk University) ;
  • Lee, Si-Kyung (Department of Applied Biology and Chemistry, Konkuk University) ;
  • Heo, Seok (Department of Food Science, University of Arkansas) ;
  • Kim, Soo-Young (Department of Biological Science, Konkuk University) ;
  • Lee, Hyung-Hoan (Department of Biological Science, Konkuk University) ;
  • Kwon, Tae-Jong (Department of Microbial Engineering, Konkuk University)
  • Published : 2004.08.01
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Abstract

A fibrinolytic enzyme has been found in several bacteria isolated from fermented food. This study was carried out to investigate the purification and characteristics of the fibrinolytic enzyme produced by Bacillus subtilis KCK-7 originated from Chungkookjang. The fibrinolytic enzyme was purified to homogeneity from the culture supernatant using ammonium sulfate fractionation and chromatographies on DEAE-cellulose and on Sephadex G-100. The final specific activity of the purified enzyme increased 11.0-fold, and the protein amount in the purified enzyme was about 16% of that in the culture supernatant. The molecular weight of the purified enzyme was estimated to be about 45,000 by SDS-PAGE. The optimum pH and temperature for the enzyme activity were pH 7.0 and $60^{\circ}C$, respectively. The enzyme activity was relatively stable up to $60^{\circ}C$ over the pH range of 7.0-10.0. The fibrinolytic enzyme activity increased by $Ca^{2+}$ and $Cu^{2+}$, whereas it was inhibited by $Hg^{2+}$ and $Ba^{2+}$. In addition, it was severely inhibited by PMSF and DFT. It is suggested that the purified enzyme was a serine protease for the fibrinolysis. The purified enzyme could completely hydrolyze fibrin in vitro within 8 h. Hence, it is suggested that the purified enzyme can be put into practice as an effective thrombolytic agent.

Keywords

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