AbstractThe goal of the present study was to investigate production of cellulase in low cost medium by thermotolerant yeast. After screening, an efficient yeast isolate having capability of C1 (exo-gluconase) and Cx (endo-gluconase) production was isolated and designated as strain R-1. Maximum enzyme production was achieved at 50 °C, pH 5.5 in the medium containing bagasse powder 4% (w/v), and ammonium sulphate 0.1% (w/v) after 72 hours of incubation. The composition containing bagasse powder, 4% (w/v); ammonium sulphate, 0.5 % (w/v); and glucose, 0.5% (w/v) achieved better production after complete medium optimization. The yeast isolate was able to tolerate wide ranges of temperature, pH, and substrate concentration for higher enzyme production. The isolated yeast was able to produce C1 (exo-gluconase) and Cx (endo-gluconase) enzymes in appropriate concentrations on a crude cellulosic substrate. Therefore, yeast may be used to power alcohol production.