Ethanol is one of the significant inhibitors of yeast cells during fermentation. Ethanol tolerance is an important characteristic that leads to an increase in ethanol productivity and yield. Four ethanologenic ethanol-tolerant yeast strains, Saccharomyces cerevisiae (ATKU132), Issatchenkia orientalis (ATKU5-60 and ATKU5-70) and Saccharomycodes ludwigii (ATKU47) were isolated by an enrichment technique in yeast extract peptone dextrose (YPD) medium supplemented with 10% (v/v) ethanol at 30C. Among non-Saccharomyces yeasts, Sd. ludwigii ATKU47 exhibited the best ethanol-tolerance and ethanol production, which was similar to that of Saccharomyces yeast, S. cerevisiae ATKU132. The maximum range of ethanol concentrations, productivity and theoretical yield by S. cerevisiae ATKU132 and Sd. ludwigii ATKU47 were 9.46-9.82% (w/v), 1.31-1.64 g/l/h and 92.56-96.05%, respectively, at 37C from an initial D-glucose concentration of 20% (w/v). In a study of ethanol production from 28% (w/v) sugarcane molasses at 37C, these two yeasts also produced high ethanol concentrations, productivity and theoretical yields in the ranges of 8.07-8.32% (w/v), 1.12-1.15 g/l/h and 92.67-95.54%, respectively.