The current industrialization and global movement is one of the most energy-intensive era with steadily rising energy consumptions and misbalancing the supply and demand means of fossil fuels. The current global energy picture perceives biofuels as potentially viable liquid fuels. Bioethanol is comparatively a sustainable alternative to petroleum products because of environment friendly burning and a renewable alternate source of fuel. Biomass from lignocellulosic plants and food crops has been extensively researched as an alternative feedstock for the generation of biofuels. Therefore, the objective of this research work was to produce bioethanol in the bioenergy lab, Department of Energy Systems Engineering, using local plant material (hemp), fruit waste (watermelon), and sugarcane through anaerobic fermentation. The production characteristics of bioethanol were also determined to identify the effect on volumetric efficiency of bioethanol production. The feedstock pH in the mash tank was adjusted using acidic and basic contents as per requirement. The substrate mixture was then delivered to the fermentation chamber for hydrolysis and biological transformation using yeast content to convert glucose content into alcohol. The hot water circulating system was used to maintain temperature during fermentation for 72 h. The bioethanol quantity was determined from the concentrated liquid extracted from distillation process. The maximum results were observed in treatment T-8 (pH 5 and temperature 35 degrees C) for condense liquid extracted was 687 ml per unit feedstock and 5.5% ethanol production followed by T-9 with ethanol percentage of 5.0% and extracted liquid (709 ml), while the lowest ethanol production was observed in T-1 (1.3%). This research study proved the biochemical transformation of agricultural biomass into bioethanol production a feasible and environment friendly technique. Still, more detailed studies are required to get improved biological treatment of biomass for enhanced quality bioethanol production.