Modern trends of microbial biotechnologies in food industry envisage development of new ferments based on pure microbial cultures with pre-defined properties and characteristics showing favorable effect on human health. Bacteria displaying elevated growth rate and acid generation activity, producing antimicrobial agents, aromatic compounds, polysaccharides, vitamins, enzymes and other bioactive substances are engaged in formulation of such preparation. The studies on species composition of microflora from diverse milk products, screening of active microbial variants, elaboration of special starter cultures comprising industrial strains have attained global relevance. Isolation of lactic acid bacteria is carried out from various sources – self-fermented dairy products, plants, vegetables, fruits, faeces of sucklings, etc. Aim of this research was recovery of lactic acid bacteria from fresh cow milk and their physiological-biochemical characterization. The experiments were performed using methods of submerged microbial culture in liquid, agar and semi-solid nutrient media, spectrophotometry, microscopy, analytical techniques to evaluate acidogenic potential, assays of enzyme activity. As a result 29 cultures were isolated and affiliated to lactic acid bacteria in accordance with their cultural-morphological and biochemical properties. 4 isolates are distinguished by maximum growth rate at 43°C, the others are mesophiles with grows optimum range 28 – 37°C. Correlations of grows and acid generation under optimal conditions were investigated in tested lactobacteria. It was found that by 24h of fermentation on MRS medium the level of biomass accumulation differed two or even three-fold among bacterial strains, the titratable acidity varied from 46 to 200°T, pH value ranged from 4,2 to 6,7. NaCl resistance is a vital technological parameter of microbial cultures applied in food industry. It was shown that biomass accumulation by isolates in the presence of 4% NaCl in the nutrient medium reached 70–100% of the control level, whereas build-up of salt concentration to 6,5% decreased biomass accumulation by 40–80%. Examination of antagonistic activity by wells technique has revealed 6 isolates suppressing growth of Salmonella typhimurium and 3 antagonists of Staphylococcus aureus. In liquid nutrient media 6 isolates inhibited growth of Staphylococcus aureus by 100%, 5 strains restrained growth of Salmonella typhimurium by 40–70%. Significant distinctions were established in caseinolytic activity of tested cultures. Upon 7 days bacterial growth on skim milk media free aminoacid content (calculated as tyrosine) in 15 strains averaged 0,3–0,5 μM/ml. 14 strains expressed enhanced activity so that free aminoacid ratio rose up to 3,2–3,4 μM/ml. Identification of the bacteria using Bruker Daltonik MALDI Biotyper system allowed to refer 90% of isolates to species Enterococcus faecalis. The completed studies resulted in 7 strains of bacteria Enterococcus faecalis distinguished by high growth rate, intense acidogenesis, NaCl tolerance, proteolytic activity, antagonistic potential, production of bacteriocin-like compounds.