Growth, antibacterial and proteolytic activities of two new lactobacilli strains isolated from matsoun (Armenian traditional dairy product) and their pH dependence were studied. The results demonstrated the antibacterial activity of lactobacilli against Gram‐positive and Gram‐negative test strains. This activity was stable when pH of cell culture medium was adjusted to the value of 6.5. At pH 8, the antibacterial activity of only one strain was stable. Both strains were able to hydrolyse casein in pH range of 5.5–8 with maximal activity at pH 5.5. Controlled pH conditions were suitable for biomass yield, while noncontrolled pH was better for expression of antibacterial activity.