This research examined the possible pathway of monosaccharide production from the rice straw waste using three integrated enzymatic hydrolysis approaches: boiled hot water pre-treatment with enzyme, alkaline pre-treatment with enzyme, and acid pre-treatment with enzyme, that can be further used as the feedstock for anaerobic digestion. Two cellulase enzymes: SIGMA-ALDRICH laboratory grade cellulase from Aspergillus niger and atres Zymix plus as a commercial cellulase enzyme were applied. It was found that the boiled hot water pre-treatment with the commercial cellulase gave the highest total monosaccharides yields. Glucose was the most significant part (78–86%) of the monosaccharides. For the pre-treatment with dilute acid, glucose was also the main component of monosaccharides; however, for the alkali pre-treatment, xylose was the main monosaccharide. It made up 48–85% of the total monosaccharide compared to glucose that made up 5–49% of total monosaccharide. Boiled rice straw with commercial cellulase enzyme provided the highest glucose yield compared to other experiments. Moreover, the obtained results from GC–MS/MS analysis show that up to 62 species of phenolic compound could be found in enzymatic hydrolysis of the rice straw waste. Aromatic and aliphatic hydrocarbon substances were also detected in the FEEM analysis. From the overall results, the integrated enzymatic hydrolysis with boil hot water pre-treatment was the most efficient method for monosaccharide production from the rice straw waste.