Simultaneous saccharification and fermentation was similarly conducted to the procedure described in NREL/TP-510-42630 [33]. Briefly, the pretreated Miscanthus containing 3-9 wt% glucan was load into a 250 ml flask and 10 ml og 1 M citrate buffer (pH 4.8), 20 FPU cellulase/g cellulose (Novozyme, cellic Ctec ll), 10 ml of YP (final concentration; peptone 20 g/L, yeast extract 10 g/L) and sterilized H2O were added to prepared 90 mL of the fermentation mixture. Simultaneously, yeast cells were harvested by centrifuging at 15,00 rpm foe 10 min at 4 C. Harvested cells were washed with sterilized water, and cell pellets were obtained by centrifugation. The cells pellets were re-suspended in 10 mL of sterilized water. The cells were then added to prepared 90 mL fermentation mixture, resulting in a working volume of 100 mL. Finally, SSF was performed at 42 C for 72 h with an agitation of 150 rpm. Them, samples were taken periodically to determine the ethanol and sugar concentration using GC and HPLC, respectively.