Purification and Some Properties of Chitinase from the Stomach of Japanese Eel, Anguilla japonica

Abstract

Chitinase (EC 3.2.1.14) was purified from the stomach of Japanese eel, Anguilla japonica, by fractionations with ammonium sulfate, Sephadex G-100 gel filtration, and DEAE-cellulose, CMcellulose, and hydroxylapatite column chromatography. The molecular weight of this enzyme was 50, 000 by SDS-PAGE, and the optimum pH was 4.4. The activity was strongly inhibited by Hg²⁺ and slightly activated by EDTA. The hydrolysis products of colloidal chitin by the enzyme were GlcNAc and GlcNAc₂. When GlcNAc_3-6 was used as substrate, GlcNAc and GlcNAc₂ were recognized as final products with various ratios. GlcNAc₂ was not hydrolyzed by this chitinase.