title: Purification and characterization of chitinase from Streptomyces sp. M-20.
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 degrees C. The enzyme was stable from pH 4 to 8, and up to 40 degrees 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.
authors.affiliation: Department of Life Science, Soonchunhyang University, Asan-si 336-745, Korea. kyoungjakim@hotmail.com
authors.forename: Kyoung-Ja
authors.initials: KJ
authors.lastname: Kim
authors.forename: Yong-Joon
authors.initials: YJ
authors.lastname: Yang
authors.forename: Jong-Gi
authors.initials: JG
authors.lastname: Kim
mesh_terms: D000818:Animals; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D002688:Chitinases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000737:chemistry / Q000494:pharmacology; D007700:Kinetics; D008670:Metals / Q000737:chemistry / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D000935:Antifungal Agents; D007477:Ions; D008670:Metals; D002688:Chitinases