An enzymatic mixture of cellulases and xylanases was produced byPleurotus ostreatususing microcrystalline cellulose as inducer, partially characterized and tested in the statistical analysis ofArundo donaxbioconversion. The Plackett-Burman screening design was applied to identify the most significant parameters for the enzymatic hydrolysis of pretreatedA. donax. As the most significant influence during the enzymatic hydrolysis ofA. donaxwas exercised by the temperature (°C), pH, and time, the combined effect of these factors in the bioconversion byP. ostreatuscellulase and xylanase was analyzed by a 33factorial experimental design. It is worth noting that the best result of 480.10 mg of sugars/gds, obtained at 45°C, pH 3.5, and 96 hours of incubation, was significant also when compared with the results previously reached by process optimization with commercial enzymes.