The alcohol fermentation using glucose and lactose was carried out to study the effect of fermentation temperature on the ethanol tolerance of Saccharomyces cerevisiae STV89 and Kluyveromyces fragilis CBS397. The maximum specific growth rate and ethanol production rate were increased up to 35$^{circ}C$ with the fermentation temperature, although maximum ethanol and cell concentration were decreased by increasing the fermentation temperature. The cell viability was also improved by lowering the fermentation temperature. Under the experimental conditions, the best ethanol tolerance of yeast strains was obtain at $25^{circ}C$. The ethanol tolerance of S. cerevisiae is better than that of K. fragilis at the same fermentation condition. With respect to the carbon source, glucose is found to be more favorable for ethanol tolerance of K. fragilis than lactos.