Purification and characterization of chitinase from Gliocladium catenulatum strain HL-1-1

Chitinase was isolated and purified from Gliocladium catenulatum through ammonium sulfate sedimentation, sephadex G-25 gel filtration, deionization, ultra filtration condensation, negative ion exchange separation, and non-denaturing gel electrophoresis. A 51 kDa chitinase was purified from the fungus through sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum temperature and pH for chitinase activity were 60 degrees C and pH 6.0, respectively. The enzyme solution was stable from 20 to 40 degrees C and pH 4 to 5. Co2+ and Ca2+ were conducive to the enzyme activity, whereas the Fe3+, Cu2+, and Ag+ evidently inhibited the enzyme activity. Fe2+, Na+, K+, Zn2+, and Mn2+ showed less inhibitory effects on the enzyme activity. The K-m of the chitinase was 2.832 mg ml(-1). The chitanase were found to inhibit the hyphal growth, conidial germination and sclerotial germination of various plant pathogenic fungi.