Rapid industrialization, increasing energy demand, and climate change are the conditions that forced the researchers to develop a clean, efficient, renewable, and sustainable source of energy which has a potential to replace fossil fuels. Ethanol is one of the attractive and suitable renewable energy resources. In present study, effectiveness of microwave pretreatment in combination with sodium hydroxide (NaOH) for increasing enzymatic hydrolysis of rice straw has been investigated and under optimum conditions obtained a maximum reducing sugar (1334.79 g/mL) through microwave assisted NaOH pretreatment. Chemical composition analysis and scanning electron microscope (SEM) images showed that the removal of lignin, hemicellulose, and silicon content is more in microwave assisted NaOH pretreatment than the blank sample. X-ray diffraction (XRD) analysis revealed that the crystallinity index of rice straw treated with microwave assisted alkali (54.55%) is significantly high as compared to the blank (49.07%). Hence, the present study proves that microwave assisted alkali pretreatment can effectively enhance enzymatic digestibility of rice straw and it is feasible to convert rice straw for bioethanol production.