The growing global demand for sustainable energy solutions has led to a surge of interest in bioethanol production from lignocellulosic biomass. However, low productivity, inefficient substrate conversion, and inadequate fermentation kinetics restricts its economic. This research addresses these issues by exploring the use of nanotechnology, mainly iron oxide (Fe₃O₄) nanoparticles, to enhance fermentation efficiency for bioethanol synthesis from sugarcane bagasse. Methodologically, 100 g of sugarcane bagasse was pre-treated with 10% (w/v) sulfuric acid, followed by pH adjustment using calcium hydroxide. The hydrolyzed mixture underwent fermentation with Saccharomyces cerevisiae in the presence of varying concentrations of Fe₃O₄ nanoparticles (0.05 g, 0.10 g, 0.15 g, and 0.20 g). Bioethanol yield was monitored over 7 days, and the samples were distilled to quantify bioethanol production. Results indicate that the addition of Fe₃O₄ nanoparticles significantly improved bioethanol yield. The sample containing 0.20 g of nanoparticles achieved the highest bioethanol concentration of 19.4% (v/v) and exhibited the most substantial reduction in sugar levels from 8.5% (w/v) to 4.20% (w/v) as well as a decrease in pH from 5.50 to 3.50 over the fermentation period. ANOVA confirmed that iron oxide nanoparticles and fermentation time had a significant impact on ethanol yield (p < 0.05). Tukey HSD showed significant improvement up to 120 h with stabilization thereafter. Increased concentrations of nanoparticles had a greater impact on ethanol yield, and this reveals synergistic role with fermentation time to maximize output. Hence, the incorporation of Fe₃O₄ nanoparticles enhances the bioethanol production process from sugarcane bagasse, making it a viable method for improving bioethanol yield in lignocellulosic biomass fermentation.