Purification and some properties of chitinase from the stomach of red sea bream Pagrus major.

Abstract

Chitinase was purified from the stomach of red sea bream Pagrus major by fractionations with ammonium sulfate, Sephadex G-100 gel filtration, DEAF-cellulose, CM-cellulose and hydro-xylapatite column chromatography. The purified enzyme showed a single band on disc and SDS polyacrylamide gel electrophoresis and the molecular weight was estimated to be 46, 000. The isoelectric point was 8.3. The optimum temperature and pH were 50°C and 5.5, rerpectively. The activity was completely inhibited by Hg2+, strongly inhibited by Fe2+ and Sn2+, and slightly. activated by Cu2+. The products of hydrolysis of chitin with the enzyme were N-acetylglucosamine and N, N'-diacetylchitobiose.