Figure 7.

Specificity of the Cation-Dependent Debranching Activity toward Different Polysaccharides.

Both purified DBE activity (black triangle) and commercial isoamylase produced by Pseudomonas sp (megazyme; black circle) were incubated with 0.5% of amylopectin (Ap), glycogen (Gly), phytoglycogen (Phy), β-limit dextrin of amylopectin (BLD), phosphorylase-limit dextrin of glycogen (PL-gly), phosphorylase-limit dextrin of amylopectin (PL-Ap), semiamylopectin of the wild-type CLg1 strain (Ap-CLg1), water-soluble polysaccharide of class A mutant (WSP-X), and water polysaccharide of wild-type CLg1 strain (WSP-WT). The release of reducing ends was determined at 10, 20, 30, and 60 min of incubation using the 3,5-dinitro-2-hydroxybenzoic acid method. In order to compare both DBE activities, the specific activity of commercial isoamylase was adjusted at 0.5 μmol of reducing ends min⁻¹ mL⁻¹ using amylopectin as substrate. Data are the means ± sd of triplicates of two independent purification of cation-dependent debranching activity.