title: Purification and characterization of chitinase B from moderately thermophilic bacterium Ralstonia sp. A-471.
abstract: Chitinase B was purified from a culture medium of Ralstonia sp. A-471 by precipitation with (NH4)2SO4 and column chromatography with DEAE-Toyopearl 650 M and Sephacryl S-200. The purified enzyme was homogeneous on SDS-PAGE. The molecular weight was 45,000 by SDS-PAGE. The optimum pH was 5.0 and stable pH was from 5.0 to 10.0. In the early stage of the reaction, chitinase B produced beta-anomer of (GlcNAc)2 from the substrate (GlcNAc)6, whereas (GlcNAc)4 produced almost at equilibrium, indicating that the enzyme predominantly hydrolyzes the second glycosidic linkage from the nonreducing end of (GlcNAc)6.
authors.affiliation: Laboratory of Biocycle Engineering, Department of Applied Biological Chemistry, Osaka Prefecture University, Osaka 599-8531, Japan. mueda@biochem.osakafu-u.ac.jp
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Ueda
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kotani
authors.forename: Aji
authors.initials: A
authors.lastname: Sutrisno
authors.forename: Masami
authors.initials: M
authors.lastname: Nakazawa
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Miyatake
mesh_terms: D002688:Chitinases / Q000302:isolation & purification* / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D043367:Ralstonia / Q000201:enzymology*; D013997:Time Factors
chemical_list: D002688:Chitinases