Purification and Characterization of Chitinase from Streptomyces sp. M-20

Kyoung-Ja Kim, Yong-Joon Yang, Jong-Gi Kim
2003 BMB Reports  
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 ionexchange 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 sulfatepolyacrylamide gel electrophoresis, and was confirmed by activity staining with Calcofluor White M2R. Chitinase was optimally
more » ... was optimally active at pH of 5.0 and at 30 o C. The enzyme was stable from pH 4 to 8, and up to 40 o 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.
doi:10.5483/bmbrep.2003.36.2.185 pmid:12689517 fatcat:nofmoior3vgo5gu35uen3e627m