The choice of pretreatment is a very important in bioethanol production from lignocellulosic biomass. This helps to eliminate lignin partition between cellulose and hemicellulose. However, various methods generate diverse effects on the material structure and composition. The purpose of this study, therefore, was to delignify sugarcane bagasse by ozonolysis, followed by hydrolysis and fermentation. Also, the morphology of the samples was analyzed using SEM, while hemicellulose, cellulose, and lignin content were characterized by the Chesson process. The sample was hydrolyzed using 1% (v/v) sulfuric acid and the bioethanol fermented with Saccharomyces cerevisiae was detected by gas chromatography. Furthermore, ozone was applied for 90 minutes at pH 3.0 in the delignification process. This produces cellulose, hemicellulose, and lignin estimated at 59%, 22%, and 6%, respectively. However, ozonolysis was employed to reduce lignin for up to 217%. Meanwhile, the hydrolysed samples were known to rapidly decrease the reducing sugar from 19.342 to 2.86 mg/L after heating at 100ºC. Subsequently, the fermentation stage recorded the highest ethanol production, estimated at 0.79% (v/v). The result showed lignin removal was conducted in an eco-friendly and efficient condition. Therefore, the need for further study is possible in order to optimize certain parameters for maximum bioethanol production.