2.4.2. Enzymatic Hydrolysis

CCM hydrolysis tests were conducted in 50 mL vessels incubated at 50 ◦C and 250 rpm on an Innova 40 shaker (Edison, NJ, USA) for 48 h. A reaction mixture of 20 mL total contained 100 g/L of CCM dry matter in 0.1 M of Na-acetate buffer at pH 5.0 with 10 mg/g of dry matter protein loading of B151 enzyme preparation. To overcome the cellobiose inhibition effect, an amount (40 U/g dry substrate) of cellobiase activity of F10 enzyme preparation was added to the mixture. Then, 1 mM of NaN<sup>3</sup> and 100 µg/mL of ampicillin were used to prevent contamination. Nelson-Somogyi assay [29] was used to determine the concentration of reducing sugars in the reaction mixture.

CCM enzymatic convertibility was defined as a degree of conversion (48 h) to reducing sugars (in glucose equivalent) as a percentage per initial concentration of substrate (*w*/*w*).
