Two novel two-step pretreatments for enzymatic hydrolysis of rice hull (RH) were proposed to lower the severity requirement of fungal pretreatment time. They consisted of a mild physical or chemical step (ultrasonic and H(2)O(2)) and a subsequent biological treatment (Pleurotus ostreatus). The combined pretreatments led to significant increases of the lignin degradation than those of one step pretreatments. After enzymatic hydrolysis of the pretreated RH, the net yields of total soluble sugar (TS) and glucose (G) increased greatly. The combined pretreatment of H(2)O(2) (2%, 48 h) and P. ostreatus (18 d) was more effective than sole pretreatment of P. ostreatus for 60 d. It could remarkably shorten the residence time and reduce the losses of carbohydrates. Ligninase analyses and SEM observations indicated that the enhancing of the efficiency could possibly attribute to the structure disruption of the RH during the first pretreatment step. So, the combined pretreatment could be recommended to different lignocellulosic materials for enzyme based conversions.