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Oil palm empty fruit bunch (OPEFB) constitutes a great source of lignocellulosic biomass, mainly comprising of 66.97% of holocellulose (cellulose and hemicellulose) and 24.45% of lignin. This present work aimed to hydrolyze cellulose present in OPEFB to form glucose with aid of Aspergillus niger. Aspergillus niger is a type of filamentous fungi able to produce cellulase, a multi-enzyme complex consisting of endoglucanase, exoglucanase, and -glucosidase, able to converting cellulose into glucose. The glucose produced is then fermented to produce bioethanol. In this present work, we compared hydrolytic activity of cellulose between OPEFB with preteatment using NaOH 10% and OPEFB without pretreatment, with regard to temperature, pH, and hydrolysis time. The concentration of reducing sugar derived from cellulosic hydrolysis was determined by using glucose assay of 3,5-dinitrosalicylic acid. The results showed that hydrolysis of cellulose operated at 40°C for 3 days yielded reducing sugar of 13.01 mg/mL (OPEFB-untreated) and 1.16 mg/mL (OPEFB-pretreated).