AbstractThis is the first report of the potential of Acacia fast growing trees in Thailand, A. mangium and the Acacia hybrid (A. mangium x A. auriculiformis), as raw material for ethanol production through a simultaneous saccharification and fermentation process by Saccharomyces cerevisiae TISTR 5339. Alkaline pulping was applied as the pretreatment process. Optimization of ethanol production was studied using response surface methodology based on central composite design. The optimized conditions of 100 g/L solid loading and an A600 of S. cerevisiae TISTR 5339 of 2 gave observed values of ethanol production of 35.7 and 27.3 g/L, which corresponded with the predicted values of 32.32 and 26.37g/L from A. mangium and A. hybrid, respectively. This condition was then used for up-scaling in a 10-L stirred bioreactor. The improved maximum ethanol concentrations of 37.84 and 36.52 g/L were obtained from A. mangium and Acacia hybrid, respectively, within 96 h of cultivation at 30 °C and no aeration rate.