Optimization of initial total sugar concentration of sweet sorghum juice, aeration time and aeration rate on ethanol production was performed by response surface methodology (RSM). The optimum conditions for ethanol production from concentrated sweet sorghum juice were determined as follows: initial total sugar concentration, 21.2 Brix; aeration time, 7.66h; aeration rate, 1.22 vvm. At the optimum conditions, the maximum ethanol yield was predicted to be 91.65% by model prediction. Similarly, 92.98% of ethanol yield was obtained by verification experiment using optimum conditions after 48 h of fermentation. This result was in agreement with the model prediction.