For SHF processing of enzymatic hydrolysate, 3 mL of the pre-cultured S. cerevisiae was inoculated
with 200 mL of the enzymatic hydrolysate containing reducing sugars in a 500 mL culture flask at 30 °C and 180 rpm for 20 h. SHF processing was also conducted using dilute acid hydrolysis. The acid hydrolysate was collected via centrifugation (7000 rpm for 20 min) and the pH of the hydrolysate was then adjusted to 6.0 with a 4 N NaOH solution to reach a suitable pH range for ethanol fermentation with S. cerevisiae. The fermentable sugar consumption and amount of produced ethanol were determined periodically.