in a 500 mL baffle flask and incuba

in a 500 mL baffle flask and incubated with shaking at 200 rpm and 25 C for 5 days. The supernatants were harvested after centrifugation at 7000g and 4 C for 20 min and subjected to a Bradford assay (Bradford, 1976) to measure the protein concentrations.
Reducing sugars released after hydrolysis were measured to determine the enzyme activities on 1% Avicel, 1% CMC, 1% b-glucan, 1% xylan and 0.1% pectin as substrate using 3,5-dinitrosalicylic acid (DNS) reagent and a glucose standard curve (Miller et al., 1960).