Bioethanol synthesis techniques have been studied intensively due to the energy crisis and various environmental concerns. A two-step bioethanol production process was carried out multiple times in an unbaffled agitator tank. The parameters varied, including the fermentation temperature, the pH level, the amount of yeast, and the impeller type. Then, a simulation was used to obtain an image of the agitation behavior inside the agitator tank to compare the velocity profile of each type of impeller design. The impeller with eight blades was found to produce the highest flow velocity: 0.28 m/s. The highest concentration of bioethanol generated from the fermentation was 34 g/L, which was produced by using an eight-blade impeller at 30 °C, a pH level of 5, an agitation speed of 70 rpm, and 2 wt % yeast. The two-blade impeller produced the lowest bioethanol concentration, 18 g/L, under the same conditions. Ethanol concentration was found to peak at 40 °C and a pH level of 5. The geometry of the impeller, the fermentation temperature, and the pH level were each found to have a significant effect on the resulting bioethanol concentration according to the results of an ANOVA test. The amount of yeast had no effect on the fermentation reaction. Finally, the results demonstrated the possibility of using computational fluid dynamic modeling to determine the impeller’s behavior for the development of the bioethanol fermentation process. The simulation and experimental results from this research support the scaling up of a bioethanol production facility. @en