In this study, the repeated-batch fermentation of liquefied cassava medium using the flocculent hybrid Saccharomyces cerevisiae CHFY0321 was investigated for semicontinuous, high-throughput production of bioethanol. Cassava medium was selected due to the industrial requirement for a cheap starchy substrate. Fermentations were performed by simultaneous saccharification and fermentation (SSF) with a set of ten batches successfully completing a series within the repeated fermentation process. In addition, pH of the culture medium was not controlled to simplify ethanol production for future use in industry. Optimal recycling volume was found to be 5%. Volumetric productivity, final ethanol concentration, and ethanol yield were measured at 3.34 g l(-1) h(-1), 84.5 g l(-1), and 90.7%, respectively. Cell recycling (24.5 g DCW l(-1)) resulted in 1.8-fold decrease in fermentation time (24 h) and 1.8-fold increase in volumetric productivity compared with the ordinary batch fermentation. Therefore, repeated-batch SSF using flocculent CHFY0321 demonstrates the possibility of cost-effective bioethanol production by eliminating additional saccharification and inoculation steps.