In this study, 14 yeast cultures from 62 isolates from traditional sourdoughs collected from 6 different regions of Turkey were selected by FT-IR identification and characterised to reveal their probiotic properties. Four yeast strains were genotypically identified and compared with FT-IR identification. In all analyses, it was observed that mostly Saccaromyces cerevisiae strain exhibited high hydrophobicity, auto-aggregation feature, and all yeast isolates in this study showed tolerance to 0.3%, even salt concentration. In addition, all yeast strains were susceptible to anti-yeasts agents, although they were resistant to all antibiotics used in the study. All selected yeast isolates exhibited high antimicrobial activity against the Staphylococcus aureus. In conclusion, this study investigated the potential probiotic properties of yeast strains isolated from sourdough.