Tej is Ethiopain traditional home made as well commercially available honey wine made from honey, gesho (Rhamnus Prenoides) and water. To date the development of starter cultures were not performed to Tej fermentation. In this study the possibility to prepare starter cultures for honey to Tej fermentation was shown. For the development of starter culture, first the yeasts were isolated from homogenized Tej sample using a specified media (potato dextrose agar and tartaric acid) cultured at 28 0C for 3-4 days. Then, five representative colonies of yeasts were characterized from 10-5-10-6 serial dilutions. The characterization methods include cell morphology; temperature tolerance, salt tolerance and sugar fermentation tests using YPD broth. The yeasts were identified to species level presumptively according to the conventional method of classification. The identified yeasts were Saccharomyces cerevisiae, Saccharomyces kluyveri, Saccharomyces daireness and Saccharomyces pastorianus. These were propagated and freeze dried. The yeasts effectiveness to Tej fermentation was tested against their PH, total titratable acidity and alcohol by volume of the products. The Mean values of PH ranged from 3.2-3.7 and the mean of the control was 3.27. Total titratable acidity ranged from 6.5-7.0 mL of 0.1N NaOH and the mean of the control was 7.0.The mean values of alcohol by volume ranged from 3.0-6.1 from 100 mL of honey. Saccharomyces cerevisiae was selected better next to the control which had lower PH (mean value 3.30) and higher titratable acidity (mean value 6.9). Significance difference between the starter cultures (p<0.05) were observed for the sensory attributes (appearance, taste, flavor, overall acceptability. Based on the sensorial evaluation Saccharomyces cerevisiae were also better selected next to the control. The findings of this study help to define and standardize the fermentation of honey to Tej.