Production of ethanol from molasses fermentation by different microbes was studied in a sugar factory. Sacharymyces (Yeast) and Trichoderma spp. are the predominant organisms contributed 24.44 and 35.59 percent population contribution. Trichoderma spp. can produce highest quantity of ethanol (0.52 ml/gm) when glucose is used as substrate followed by cellulose and xylose. Highest 0.37 ml/gm ethanol is produced during molasses fermentation by Trichoderma followed by Sacharomyses. Commercial ethanol production hence may be recommended from molasses using Trchoderma spp.