Fermented Ensete (Ensete ventricosum (Welw) Cheesman) commonly called false banana is an important nutrient source in southern part of Ethiopia. This study aimed at screening potential probiotic bacteria from a traditional fermented Ensete. A total of 12(twelve) samples were collected from local traditional fermented Ensete. Various morphological and biochemical tests were performed for screening of the potential probiotic bacteria as starter culture. Media optimization was carried out for cultivation of the potential isolate. The potential isolate was found to be gram positive, non motile, negative for (Catalase, indole). This bacterium had broad range of pH 4.0 – 8.0 and temperature of 25-40 (0c). It was tolerant to 1-4% salt concentration. High growth rate of the isolate was observed in the presence of 2 % glucose and 0.2 % peptone. The optimum temperature and pH for high biomass production was observed at 370c and pH of 7. Fermentation of Ensete with isolate increased the nutritional value and aroma of a food as well. The isolate was found to be Lactobacilli sp. CH2 which fulfills the required criteria for a probiotic such as tolerance to such as high salt, low pH, body temperature range. The result in this study is reproducible and reliable for further processing of Ensete in the local community. Molecular study should be done for characterization of this potential strain. Keywords: Lactobacilli, Prebiotic, Probiotic, Starter culture
