To develop a practical and cost-effective medium for bioethanol production from the hydrolysate of seaweed Sargassum sagamianum, we investigated the feasibility and performance of bioethanol production in CSL (corn-steep liquor)-containing medium, where yeast Pichia stipitis was used and the repeated batch was carried out in a surface-aerated fermentor. The optimal medium replacement time during the repeated operation was determined to be 36 h, and the surface aeration rates were 30 and 100 ml/min. Under these conditions, the repeated-batch operation was successfully carried out for 6 runs (216 h), in which the maximum bioethanol concentrations reached about 11-12 g/l at each batch operation. These results demonstrated that bioethanol production could be carried out repeatedly and steadily for 216 h. In these experiments, the total cumulative bioethanol production was 57.9 g and 58.0 g when the surface aeration rates were 30 ml/min and 100 ml/min, respectively. In addition, the bioethanol yields were 0.43 (about 84% of theoretical value) and 0.44 (about 86% of theoretical value) when the surface aeration rates were 30 ml/min and 100 ml/min, respectively. CSL was successfully used as a medium ingredient for the bioethanol production from the hydrolysate of seaweed Sargassum sagamianum, indicating that this medium may be practical and cost-effective for bioethanol production.