Probiotics provide several health benefits that transcend basic nutritional requirements of our daily meal. The focus of this study is to evaluate the antimicrobial capability of probiotic bacteria isolated from commercially available yoghurts in Indore city. Three cans of different branded yoghurts were purchased from local supermarkets and prepared using standard procedures. The samples were transferred into De Man, Rogosa and Sharpe (MRS) broth and incubated anaerobically for 24 hrs. The isolates were identified using biochemical tests. The antibacterial activity was evaluated using agar diffusion assay. The effects of pH and temperature on the growth of isolated Lactobacillus spp. as well as their ability to tolerate bile salt were assayed. All isolated lactobacillus spp. has inhibitory effect on E. coli, S. aureus and P. aeruginosa. Bile salts incorporated in the bacteria cultures were effective on bacterial viability. The present study also focused on the antimicrobial activity of cell free culture filtrate and extracellular protein concentrate of Lactobacillus strain against food-spoilage organisms and human pathogens. Though both bacteria and fungi that have beneficial health effects are categorized as probiotics, Lactobacillus spp. have been well characterized as prominent members of the probiotic group. The probiotic bacteria isolated in commercially available yoghurts have health beneficial effects, which include antibacterial activities.