The quest to reduce the level of overdependence on fossil fuel product and to provide all required information on proven existing alternatives for renewable energy has resulted into rapid growth of research globally to identify efficient alternative renewable energy sources and the process technologies that are sustainable and environmentally friendly. The present study is aimed at production and characterization of bioethanol produced from sugarcane juice using a 24 factorial design investigating the effect of four parameters (reaction temperature, time, concentration of bacteria used and amount of substrate). The optimum bioethanol yield of 19.3% was achieved at a reaction temperature of 30oC, time of 72 hours, yeast concentration of 2 g and 300 g concentration of substrate (sugarcane juice). The result of statistical analysis of variance shows that the concentration of yeast had the highest effect of 7.325 and% contribution of 82.72% while the substrate concentration had the lowest effect and% contribution of-0.25 and 0.096% respectively. The bioethanol produced was then characterized for some fuel properties such as flash point, specific gravity, cloud point, pour point, sulphur content, acidity, density and kinematic viscosity. The results of bioethanol characterization conform to American society for testing and materials (ASTM) standard. Hence, sugarcane juice is a good and sustainable feedstock for bioethanol production in Nigeria owing relative abundance, cheap source of supply and available land for large scale production.