An efficient pretreatment with various concentrations of FeCl3 (0.005-0.2 mol/L) was developed to extract hemicellulose in sugarcane bagasse and enhance the enzymatic hydrolysis of cellulose in pretreated solids. It was found that 0.025 mol/L FeCl3 pretreated substrate yielded a high glucose yield of 80.1% during enzymatic hydrolysis. Then the characterization of raw material and pretreated solids was carried out to better understand how hemicellulose removal affected subsequent enzymatic hydrolysis. In addition, Tween 80 and Bovine Serum Albumin (BSA) were added to promote enzymatic hydrolysis of pretreated substrate. Together with that obtained from pretreatment, the highest glucose yield reached 97.7% with addition of Tween 80, meanwhile, a reduction of 50% loading of enzyme yielded the same level of glucose. However, the increased yields with additives decreased gradually as the hydrolysis time was extended. Furthermore, the enhancement mechanisms of Tween 80 and BSA were determined.