Introduction: Probiotics are defined as living microorganisms, when ingested in sufficient amounts provide health benefits for the host. The two main types of probiotics are Lactobacillus and Bifidobacterium. Probiotics can be used in the form of powders, syrups, tablets and fortified foods. Application of probiotics and prebiotics as biotherapeutics is the new field in developing dietary strategies. Materials and Methods: To identify the isolated bacteria, microbial and biochemical tests including gram staining, catalase and oxidase activity, sugar fermentation of carbohydrates, antibacterial tests and acid and bile salts resistance were performed. The asparaginase and glutaminase production was evaluated by raid plate assay. Also, bacterial DNA was extracted and 16S rRNA gene was amplified through PCR using universal primers. Results: The results confirmed the isolated bacteria as a probiotic including; gram-positive, catalase- and oxidase-negative, grow at bile salts and acidic pH and no hemolysis. Based on 16S rRNA gene amplification, it was identified as Lactobacillus. Furthermore, the production of therapeutic enzymes asparaginase and glutaminase was verified. Conclusion: The isolated bacteria from traditional buttermilk confirmed as lactobacillus and further experiments about asparaginase and glutaminase production are running.
