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An Indian traditional fermented food, idli batter, was used as a source for isolation of lactic acid bacteria (LAB). A total of 15 LAB strains were isolated on the basis of their Gram nature and catalase activity. Of these, one lactobacilli strain and one lactococci strain which showed antimicrobial activity were identified using biochemical characterization, sugar utilization and molecular sequencing. The microbes, labeled as IB-1 (Lactobacillus plantarum) and IB-2 (Lactococcus lactis) were tested for their in vitro tolerance to bile salts, resistance to low pH values and acidifying activity. Both the strains showed good viability (IB1- 58.11%; IB2- 60.84%) when exposed to high bile salt concentration (2%) and acidic pH of ≤pH 3.0 (IB1- 88.13%; IB2- 89.85%). Lactic acid (IB1- 181.93 mM; IB2- 154.44 mM), biomass production (IB1- 0.65; IB2- 0.58 g/l) after 54 h as well as qualitative estimation of β-galactosidase and vitamin B12 production were also studied to check their suitability as an industrially important strain for production of important biomolecules.