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The reduction in the operational cost of a biofuel refinery is vitally important to make biofuel competitive with fossil fuels. The aim of this paper is to find a cost-efficient and sustainable refinery capacity for grain-based ethanol (i.e., corn-based ethanol) production, which will play an important role in promoting the widespread adoption and sustainable use of ethanol, by improving the productivity of the overall refining process. Continuous-event simulation was utilized in this study to model complex operations of a refinery such as the loading, unloading and treatment of feedstock over nine major phases (e.g., feedstock storage and handling, pretreatment and conditioning, fermentation and hydrolysis, and enzyme production) to produce ethanol. To improve the model prediction, the real data of corn yield produced in Tazewell County, Illinois, U.S. were used. The proposed simulation model is implemented in AnyLogic® University 8.6.0 simulation software, Chicago, IL, USA, and the (near) optimal number of reactors for the hydrolysis and fermentation is found via optimization software known as OptQuest®, Boulder, CO, USA, As a result, the proposed approach found that six reactors showed the optimal daily profit from USD 67,500 to 82,217. This information will help engineers and policy makers to modify the capacity of a biofuel refinery for enhancement of the system efficiency and ethanol production. [ABSTRACT FROM AUTHOR]