The pretreated pulp slurries were enzymatically hydrolyzed to
determine the maximum obtainable sugar. The pH of the slurries
was preadjusted to 5.0 with sodium hydroxide, and 3 M sodium
acetate buffer at pH 5.0 was added at a final concentration of
50 mM. Cellulasewas used in the ratio of 3 U/g dry pulp. Hydrolysis
of cellulosic fiber in the slurries was conducted at 50 C for 72 h
before subsequent starch hydrolysis. Samples were collected after
24, 48, and 72 h for analysis of released glucose. After the slurrywas
treated with cellulase, a-amylase (300 U/g dry pulp) and glucoamylase
(100 U/g dry pulp) were added to hydrolyze the starch
completely. Starch hydrolysis in the slurry was conducted at 50 C
for 48 h. The amount of released glucose was determined using the
Wako glucose CII test (Wako Pure Chemicals Co., Osaka, Japan).
Clear supernatants were obtained from the hydrolyzed slurries by
centrifugation at 7000 rpm for 20 min.