Sodium methoxide (CH3ONa) with glycerol pretreatment (CWGP) was performed to improve the enzymatic digestibility of sugarcane bagasse (SCB). Response surface methodology was utilized to optimize the CWGP parameters for pretreating SCB from the perspective of total fermentable sugar yield (TFSY) and total fermentable sugar concentration (TFSC). Under the optimal CWGP conditions, 0.5666 g/g of TFSY (0.82% (CHONa)-O-3, 1.11 h, 150 degrees C) and 17.75 g/L of TFSC (0.87% (CHONa)-O-3, 1.38 h, 149.27 degrees C) were achieved, corresponding to delignification of 79.05% and 79.34%, respectively. Compared the pretreatment using glycerol or (CHONa)-O-3 alone, the CWGP has significant synergies to enhance the enzymatic efficiency of SCB. The physical and chemical characteristics of untreated and pretreated SCBs were analyzed using FT-IR, XRD, and SEM, and the results suggest that CWGP significantly increased the susceptibility of the substrates to enzymatic digestibility. Ultimately, CWGP might be a prospective candidate for the pretreatment process of enzyme-based lignocellulosic biorefineries.