Yeast is emerging as potential probiotic organism.Already there is amarked increase in the sale of various yeast based probiotic products. However there is a need to screen more potential probiotic strains of Sacharomyces and to develop new probiotic based food products. The main emphasis of the present study is screening and evaluation of probiotic properties of the yeast cultures isolated from various food sources. In this study ten isolates of food grade yeast were isolated from various food sources and identified using morphological, physiological and biochemical characteristics. The ten isolates were screened for their thermo, osmo, acid tolerance, antimicrobial properties and ï¢- galactosidase enzyme activity.Among the ten isolates one potential isolate was selected for the fermentative production of yeast. The mediumand physical conditions were optimized. BakerÂs yeast is used as reference yeast culture throughout the study. The optimized medium resulted in increased production of yeast compared to the initial production. The Sacharomyces sp. isolated fromgrapesG4 shows maximumgrowth at optimized conditions and is having high osmo, thermo and acid tolerance as compared to bakerÂs yeast and has high antimicrobial activity as compared to the other yeast cultures isolated fromcurd sample. Based on the above results obtained Sacharomyces sp. isolated fromgrapes G4 was found to be the potential isolate and may be used for further human application that is either direct food or in the preparation of functional foods to enhance the health by improving the balance of micro flora in the intestine.