A novel antifungal chitinase from Chaetomium globosum: column-free purification and characterization

A new chitinase gene, cloned from the biocontrol Chaetomium globosum W7, was designated Cgchi18. Recombinant protein Cgchi18 with 535 amino acids was expressed in Escherichia coli, and purified by means of a column-free purification method relying on split intein, achieving a 12.39-fold purification and a 15.61% recovery yield. The maximum activity of this approximately 60-kDa protein was observed at 45 °C and pH 5.0. Cgchi18 was activated by Mg²⁺ and Ba²⁺, but inhibited by Mn²⁺, Co²⁺, Cu²⁺, Zn²⁺, Ag⁺ and Hg²⁺. Cgchi18 showed high substrate specificity, only hydrolyzing β-1,4-glycoside bond in chitin and its derivatives, to liberate disaccharides or trisaccharides. For the degradation of colloidal chitin under optimal conditions, Vmax and Km of Cgchi18 were calculated as 8.05 μmol/min/mg and 3.18 mg/mL, respectively. Additionally, it exhibited antifungal activity and could have a degrading effect on the spread of hyphae of pathogenic fungi. In conclusion, the chitinase Cgchi18 identified from C. globosum has potential for industrial and agricultural applications.