Rice straw (RS) is one of the main agriculture residues in Egypt. So, this study was performed on rice straw as a resource for production of bioethanol. Fermenting agricultural biomass, such as rice straw to ethanol is a promising solution to an ongoing waste problem. However, the biomass must first be pretreated to break down lignin, thereby increasing accessibility of the substrate to fermentative organisms. Biological pretreatment by microorganisms represents a potentially economic strategy to prepare the biomass for fermentation. In a series of laboratory experiments, rice straw was pretreated with sodium hydroxide, sulfuric acid and hydrogen peroxide in single and combination treatments followed by biological treatments for bioethanol production by bacteria; Bacillus subtilis were successfully grown on cellulose, hemicellulose of rice straw hydrolysis. Acid hydrogen peroxide treatment was found to be the best chemical treatment. Cellulases from Bacillus subtilis was produced in suitable quantities (2.2 IU ml-1) after 144 h, where Simultaneous Saccharification and Fermentation (SSF) of cellulose by Saccharomyces cerevisiae and cellulases was evaluated in basal media and ethanol was produced at 18.9% v/v (189 mg/g of dry mass) after 72 h at 30 C and pH 5. DNS, FTIR, XRD and GC analysis were done.