Carboxy-terminus truncations of Bacillus licheniformis SK-1 CHI72 with distinct substrate specificity
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Bacillus licheniformis SK-1 naturally produces chitinase 72(CHI72) with two truncation derivatives at the C-terminus, one with deletion of the chitin binding domain (ChBD), and the other with deletions of both fibronectin type III domain (FnIIID) and ChBD. We constructed deletions mutants of CHI72 with deletion of ChBD (CHI72ΔChBD) and deletions of both FnIIID and ChBD (CHI72ΔFnIIIDΔChBD), and studied their activity on soluble, amorphous and crystalline substrates. Interestingly,when equivalent amount of specific activity of each enzyme on soluble substrate was used, the product yield from CHI72-ΔChBD and CHI72ΔFnIIIDΔChBD on colloidal chitin was 2.5 and 1.6 fold higher than CHI72, respectively. In contrast,the product yield from CHI72ΔChBD and CHI72ΔFnIIID-ΔChBD on β-chitin reduced to 0.7 and 0.5 fold of CHI72,respectively. These results suggest that CHI72 can modulate its substrate specificities through truncations of the functional domains at the C-terminus, producing a mixture of enzymes with elevated efficiency of hydrolysis. [BMB reports 2011; 44(6):375-380]
Abstract
Bacillus licheniformis SK-1 naturally produces chitinase 72(CHI72) with two truncation derivatives at the C-terminus, one with deletion of the chitin binding domain (ChBD), and the other with deletions of both fibronectin type III domain (FnIIID) and ChBD. We constructed deletions mutants of CHI72 with deletion of ChBD (CHI72ΔChBD) and deletions of both FnIIID and ChBD (CHI72ΔFnIIIDΔChBD), and studied their activity on soluble, amorphous and crystalline substrates. Interestingly,when equivalent amount of specific activity of each enzyme on soluble substrate was used, the product yield from CHI72-ΔChBD and CHI72ΔFnIIIDΔChBD on colloidal chitin was 2.5 and 1.6 fold higher than CHI72, respectively. In contrast,the product yield from CHI72ΔChBD and CHI72ΔFnIIID-ΔChBD on β-chitin reduced to 0.7 and 0.5 fold of CHI72,respectively. These results suggest that CHI72 can modulate its substrate specificities through truncations of the functional domains at the C-terminus, producing a mixture of enzymes with elevated efficiency of hydrolysis. [BMB reports 2011; 44(6):375-380]
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