A corn fuel ethanol plant integrated with anaerobic digestion treatment of thin stillage increases the net energy balance. Furthermore, the anaerobic digestion effluent (ADE) can be reused as a potential substitute for process water in the ethanol fermentation. In this study, the suitability of ADE as process water for corn ethanol fermentation was investigated by analyzing the potential inhibitory components in the ADE. It was found that ammonium influenced the growth and metabolism of Saccharomyces cerevisiae. Maximum ethanol production was obtained when the concentration of ammonium nitrogen was 200 mg/L, and ammonium could replace urea as the nitrogen source for S. cerevisiae under this concentration. In the ethanol fermentation with a higher concentration of ammonium, more glycerol was produced, thereby resulting in the decrease of ethanol production. In addition, components except ammonium in the ADE caused no inhibition to ethanol production. These results suggest that ADE could be reused as process water for corn ethanol fermentation without negative effect when ammonium concentration is well controlled.