Molasses is a viscous by-product of refining sugarcane or sugar beets into sugar that has been considered as a cheap source of raw materials for industrial ethanol production. In this study, the ability of high-temperature ethanol production from molasses using thermotolerant yeast Saccharomyces cerevisiae Y81 was investigated at different temperatures (35, 37, and 40^oC) on the scales of 100 mL and 2 liters. The fermentation experiments were carried out in molasses with the initial sugar concentration of 26^oBrix, pH of 4.5 and cell concentration of 10⁷ cells/mL for 6 days of fermentation. In the fermentation volume of 100 mL, ethanol concentrations of 6.13%, 6.10% and 4.50% (v/v) were achieved at 35, 37, and 40°C, respectively. The results of the ethanol production at 37°C from 2 liters of molasses showed that the ethanol concentration of 7.80% (v/v) and fermentation efficiency of 83.40%, these levels were the highest values in comparison to the other treatments at two different temperatures. Ethanol concentrations of 6.86% and 4.84% (v/v) with the fermentation efficiencies of 63.56% and 81.54% were obtained at 35°C and 40°C, respectively. From these results, 37^oC was selected as the best temperature to carry out the fermentation experiment on the scale of 50 L with the ethanol concentration of 8.10% and the fermentation efficiency of 85.98% were achieved.