AbstractA Plackett-Burman design (PBD) combined with a steepest ascent approach is a powerful technique to screen the important operating parameters for the production of reducing sugars from sugarcane bagasse (SB). In this study, the most significant parameters (p< 0.05), as identified by PBD, were as follows: pretreatment duration, pH of pretreatment process, loading of enzyme cellulase, SB loading, and moisture content of SB. Analysis of variance (ANOVA) results showed that the model of reducing sugar productivity was able to provide a high correlation between the response and its parameters. Thus, the path of steepest ascent (PSA) method was used to assess the optimal region of variables for improved reducing sugar productivity from SB. The PSA analysis revealed that by treating 7.25 g of SB (with 84% moisture content) for 82.0 minutes at a pH of 8.8, followed by the addition of 34.0% v/w of cellulase, a reducing sugar productivity of 0.03 g/L could be achieved per hour during the enzymatic saccharification process.