A total of 14 newly isolated, indigenous yeast strains from molasses and grapes, along with the control yeasts D1–D3, were investigated for ethanol production abilities and related parameters like ethanol and sugar tolerance. Strains M1, M5, M9, M12 and M15 were found ethanol resistant up to 12 % ethanol and yeast strains G1, G2, M1, M5, M6, M11, M12 and M15 showed resistance up to 20 % (w/v) glucose concentration. Among new isolates M12 gave maximum alcohol, [9.13 % (v/v)] with pure glucose followed by M15 [8.43 % (v/v)], M1 [7.37 % (v/v)] and M6 [7.26 % (v/v)] in 72 h. Ethanol productivity, after 24 h, was found to be maximum in case of M12, M3, M6, M5 and M15, in reducing order, ranging from 1.08 to 1.37 gethanol/100 gsubstrate/h and ethanol yield after 72 h was maximum in case of M12, M15, M1 and M6 ranging from 38.15 to 47.97 gethanol/100 gsubstrate. All the newly isolated strains lag behind control strains D1–D3 in productivity (1.32–1.98 gethanol/100 gsubstrate/h) as well as in yield (47.55–47.92 gethanol/100 gsubstrate). Sugarcane molasses medium having 15–16 % fermentable sugars along with other yeast nutrients, was further tested for alcohol production at 30 °C under stationary conditions, by selected yeast strains M1, M5–M7, M12 and M15, added at 0.25 % (w/v) on wet weight basis. Except M1, M5 and M12, all the strains, including control strains, produced considerable amount of alcohol ranging from 8.12 to 9.14 % (v/v) in 48 h but, simultaneous generation of high yeast biomass calls for optimization of fermentation parameters, especially aeration and agitation, for improved ethanol production.