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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Purification and Characterization of Chitinase from Streptomyces sp. M-20

  • Kim, Kyoung-Ja (Department of Life Science, Soonchunhyang University) ;
  • Yang, Yong-Joon (Department of Horticultural Science, Sangmyung University) ;
  • Kim, Jong-Gi (Department of Horticultural Science, Jungang University)
  • Published : 2003.03.31
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Abstract

Chitinase (EC 3.2.1.14) was isolated from the culture filtrate of Streptomyces sp. M-20 and purified by ammonium sulfate precipitation, DEAE-cellulose ion-exchange chromatography, and Sephadex G-100 gel filtration. No exochitinase activity was found in the culture filtrate. The molecular mass of the purified chitinase was 20 kDa, estimated by a sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and was confirmed by activity staining with Calcofluor White M2R. Chitinase was optimally active at pH of 5.0 and at $30^{\circ}C$. The enzyme was stable from pH 4 to 8, and up to $40^{\circ}C$. Among the metals and inhibitors that were tested, the $Hg^+$, $Hg^{2+}$, and p-chloromercuribenzoic acid completely inhibited the enzyme activity. The chitinase activity was high on colloidal chitin, chitotriose, and chitooligosaccharide. The purified chitinase showed antifungal activity against Botrytis cinerea, and lysozyme activity against the cell wall of Botrytis cinerea.

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References

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