Hydrolysis of waste office paper (WOP) into fermentable sugars is an important option of WOP utilization. In this work, the effect of major chemicals in WOP on its hydrolysis using industrial cellulase Accellerase 1000 (Genencor, Rochester, NY, USA) was investigated, and calcium carbonate (CaCO3) was found to be the key parameter affecting the enzymatic hydrolysis efficiency of WOP. The pretreatment methods, acid washing and acid presoaking, were tested
for the removal of CaCO3 from WOP. It was found that the presoaking of sulfuric acid (H2SO4) in WOP was an effective way. The pretreating parameters of WOP were studied on maximizing the hydrolysis efficiency. The conversion yield of cellulose to glucose and cellobiose using the pretreated WOP reached 73.3% after 96 hours hydrolysis at the optimal conditions. The results provided the WOP utilization with a practical enzymatic hydrolysis method with industrial application potential.