Bamboo biomass was widely considered as a promising substitute for lignocellulose to produce fermentable sugars and biofuels in the south of China. When P. amarus were treated using hydrogen peroxide and acetic Acid pretreatment in the presence of sulphuric acid at 60 ℃ for 2 h, 82.63% lignin was removed from the bamboo residue, and enzymatic saccharification yield of 79.3% and ethanol content of 13.31 g/L were obtained. Analysis indicated that HPAC pretreatment increased the hydrophilic and porous nature of substrate, which can improve the enzyme accessibility to cellulose. When HPAC-pretreated D. sinicus, B. lapidea, N. affinis, and D. giganteus were used as the substrates of enzymatic saccharification, glucose yields of 71–84% at 72 h were achieved. HPAC pretreatment was a highly efficient and environmentally friendly method for bamboo biorefinery in the south of China.