In the present work, two-step pretreatments were explored in order to find an effective method to enhance the reducing sugar yield of WBY. WBY was first treated by four methods including 0.1 M NaOH treatment, high speed homogenizer, ultrasonication and microwave treatment followed by hydrolysis at 121 °C for 20 min under mild acid condition (pH 2) as a second step. The ef- fects of the four pre-treatments and the optimum pre-treatment conditions were evaluated by the reducing sugar yield after hydro- lysis. Furthermore, treated WBY hydrolysates were directly served as only nutrient source for BC production by G. hansenii CGMCC3917 and we investigated the influences of the four pre- treatments and the WBY hydrolysate concentration on the BC yield. Additionally, water holding capacity (WHC), water release rate (WRR) and water absorption rate (WAR) of the bacterial cellu- lose were measured and its microstructure was determined using scanning electron microscopy (SEM).