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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Purification and Characterization of a Collagenase from the Mackerel, Scomber japonicus

  • Park, Pyo-Jam (Department of Chemistry, Pukyong National University) ;
  • Lee, Sang-Hoon (Department of Chemistry, Pukyong National University) ;
  • Byun, Hee-Guk (Department of Chemistry, Pukyong National University) ;
  • Kim, Soo-Hyun (Department of Food Science and Engineering, Cheju National University) ;
  • Kim, Se-Kwon (Department of Chemistry, Pukyong National University)
  • Published : 2002.11.30
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Abstract

Collagenase from the internal organs of a mackerel was purified using acetone precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, gel filtration chromatography on a Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel, and gel filtration chromatography on a Sephadex G-75 column. The molecular mass of the purified enzyme was estimated to be 14.8 kDa by gel filtration and SDS-PAGE. The purification and yield were 39.5-fold and 0.1% when compared to those in the starting-crude extract. The optimum pH and temperature for the enzyme activity were around pH 7.5 and $55^{\circ}C$, respectively. The $K_m$ and $V_{max}$ of the enzyme for collagen Type I were approximately 1.1 mM and 2,343 U, respectively. The purified enzyme was strongly inhibited by $Hg^{2+}$, $Zn^{2+}$, PMSF, TLCK, and the soybean-trypsin inhibitor.

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References

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