Abstract: Bacteriocin producing lactobacilli were obtained from different sources and were previously characterized outdoors. Lactobacillus bulgaricus 761N, Lactobacillus fermentum DSMZ 20049, Lactobacillus delbrueckii subsp. bulgaricus NCTC 12197 T and Lactobacillus delbrueckii subsp. bulgaricus DSMZ 20080T were the strains under study. Subcellular fractions were investigated for their proteolytic activity that results in production of bioactive peptides in the cell-free supernatant (bacteriocin). A single strain bacteriocin was selected depending upon cytotoxicity of the strain’s bacteriocin and antibacterial capacity. The cell-free supernatants of the tested strains were tested for their antibacterial capacity against a broad spectrum of pathogens involving Staphylococcus aureus, Pseudomonas aeruginosa, Burkholderia cepacia, Salmonella spp., Escherichia coli and Shigella spp. The selected candidate was Lactobacillus bulgaricus 761N. The bacteriocin was partially purified and assured via applying the semi-purified samples on SDS-PAGE. Advance to the purification process, the fractions selected were tested for their antibacterial capacity as well. Both fractions have been found to possess antibacterial activity but the lower molecular weight was over succeeded.