Figure 5

Binding assay of the truncated enzymes toward insoluble polysaccharides. The purified protein (15-20 μg) and the indicated substrate (10 mg) were thoroughly mixed at 4°C for 1 h. A, Binding assay of the four active enzymes (PglA, rPglA∆C, rPglA∆N, rPglA-CD). The enzymatic activity against laminarin remaining in the supernatant was determined and compared to the original activity of each enzyme preparation. The binding activity was calculated by subtraction of the activity recovered from the original activity which was considered as 100%. The values represent the average of the results from triplicate experiments. B, Binding of rPglA-N and rPglA-C to insoluble carbohydrates. The amount of protein remaining in the supernatant (S) and co-precipitating with the substrate (P) were examined by SDS-PAGE.