AbstractPretreatment of sawdust using a combination of sodium hydroxide (NaOH) and monochloroacetic acid (MCA) was investigated for the formation of reducing sugars. Optimum conditions for the pretreatment process were determined by the amount of reducing sugars formed during the enzymatic hydrolysis of the pretreated substrate. It was found that mercerization by NaOH played an important role in increasing the degree of substitution (DS) and that the maximum solubility was achieved during the etherification by MCA. A maximum amount of 34.2% lignin was removed in the process. As the DS of the substrate was increased, the efficiency of the hydrolysis process increased, leading to the higher yield of reducing sugars. The optimum operating conditions for the pretreatment process were determined to be 75 ºC at 90 rpm for 4 hours (2 hours for mercerization plus 2 hours for etherification). Under these operating conditions, with 1% (w/v) NaOH and 2% (w/w) MCA loading, a maximum DS of 0.2 and a solubility of 10.3% was attained. At 75 ºC and after 48 hours of the hydrolysis process, cellulases from Aspergillus niger resulted in the production of 2.88 g/L of glucose with a yield of 62.72% reducing sugars. X-ray diffraction (XRD) revealed reduced crystallinity of the sawdust and Scanning Electron Microscopy (SEM) showed distortion of the structure after pretreatment.