The goal of this research was to release fermentable reducing sugar from lignocellulose (poplar) by hot water pretreatment, and then employed to synthesize poly(3-hydroxybutyrate-co-3-hydroxy-valerate) (PHBV) using activated sludge to replace traditional petroleum-based polymers. The orthogonal experiment was used to optimize the conditions of different pretreatment temperature, pretreatment time, enzymatic hydrolysis temperature and enzymatic hydrolysis time and the optimal condition for producing sugars was pretreated at 200 °C for 30 min and enzymatic hydrolysis at 45 °C for 3d. A maximum yield was 530.3 mg/g of reduced sugar, while the furfural and 5-HMF produced in the optimum conditions were 512.61 mg/L and 239.34 mg/L. Moreover, the effects of increasing concentration of hot water pretreated poplar hydrolysates (500-1700 mg/L) on PHBV production were investigated. Poplar hydrolysate (1700 mg/L) exhibited maximum PHBV concentration of 637.556 mg/L.