Fossil energy sources are one of the driving factors in the economic development and prosperity of the nation. The availability of fossil energy sources is currently shrinking and is expected to run out quickly, so an alternative is needed to replace it. Sugarcane bagasse is a lignocellulosic biomass which can be utilized for bioethanol production as an alternative energy source. Lignocellulosic hydrolysis with the addition of H2SO4 catalyst was carried out to obtain glucose. The addition of Fe2+ and Ca2+ ions was performed to increase the glucose concentration. Commonly, Saccharomyces cerevisiae is used to convert glucose into bioethanol using fermentation process. The objective of this study was to investigate the effect of Fe2+ and Ca2+ ions on reducing sugar concentration, and the concentration of distillate bioethanol. The effect of yeast type on the concentration of bioethanol was also discussed in this paper. The result shows that the addition of Fe2+ and Ca2+ ions in the hydrolysis process can increase the concentration of glucose produced. This is because the higher concentration of cellulose, the more substrate is converted into reducing sugars. The use of Fe2+ and Ca2+ ions in the fermentation process will increase ethanol concentration compares with fermentation without metal ions. This is because Ca2+ ions have an important role in the flocculation process of bioethanol fermentation and Fe2+ ions are important nutrients for eukaryotes, which play a role in cell oxidation-reduction reactions. The type of yeast that can produce higher concentrations of ethanol is tapai yeast. This is because tapai’s yeast has three supporting ingredients for the fermentation process, whereas baker’s yeast only contains two microorganisms.