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Alcohol-based biofuel was produced through alkaline pre-hydrolysis, enzymatic saccharification and fermentation using yellow poplar as raw material. In alkaline pre-hydrolysis, 51.1% of biomass remained as residual, which chemical composition was 80.2% of cellulose, 17.6% xylan and 2.0% of lignin. Enzymatic saccharification led to almost complete conversion of cellulose and xylan in alkaline hydrolysis residue to monosaccharide. After enzymatic saccharification, the composition of hydrolysate was 95.1 g/L of glucose and 21.4 g/L of xylose. The enzymatic hydrolysate also contained 0.5 g/L of acetic acid and 0.48 g/L of total phenolics. The furfural and 5-hydroxymethylfurfural (5-HMF) was not detected in this liquor. It means that the fermentation inhibitors such as phenolics and furans are not existed or under control. The hydrolysate from enzymatic saccharification was used for the production of ethanol by Saccharomyces cerevisiae and butanol by Clostridium acetobutylicum, and C. beijerinckii, respectively. Ethanol concentration was highest for 24h fermentation as 57.5 g/L, which was 12% higher than that of control. In the YPH fermentation by C. acetobutylicum, 18.1 g/L total ABE was produced with 6.2 g/L residual xylose (productivity 0.38 g/L h, and yield 0.42). C. beijerinckii showed a little low performance in YPH fermentation. The total ABE, productivity, and yield were 12.1 g/L, 0.25 g/L h, and 0.37, respectively.