2.3.2. Enzymatic pretreatment (Variant II)
Raw material samples (30 g dry matter basis) were slurred in ca.
150 mL of distilled water in a 300 mL conical flasks and pH of the
slurry was adjusted to 4.5 using 1 mol L1 H2SO4 solution. Then
Ceremix 6X MG preparation (1.25 g of enzyme per kilogram of
raw material dry matter) was added and the total weight of the
flask content was adjusted to 200 g with distilled water. The flasks
were sealed with rubber stoppers to avoid water evaporation and
were placed in a water bath heated to 45 C for 20 min with constant
agitation [12]. Afterwards the slurries were cooled to 35 C,
inoculated with yeast, GSHE and protease (Neutrase 0.8 L) were
added and further procedure was the same as described in the Section
2.3.1.