Bioethanol has recently attracted much attention as a sustainable and environmentally friendly alternative energy source. This study aimed to develop a potential process for bioethanol production by fed-batch fermentation using instant dry yeast. To obtain the highest cell growth, we studied the influence of the initial sugar concentrations and pH of sugarcane molasses in batch fermentation. The batch system employed three levels of sugar concentrations, viz. 10%, 15%, 20% (w/v), and two levels of pH, 5.0 and 5.5. The highest cell growth was achieved at 20% (w/v) and pH 5.5 of molasses. The fed-batch system was then performed using the best batch fermentation conditions, with a molasses concentration of 13% (w/v) which resulted in high ethanol concentration and fermentation efficiency of 15.96% and 89%, respectively.