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A Lactobacillus delbrueckii strain that produces bacteriocin was isolated from the vagina of a healthy person. A multitude of bacterial species, including Pseudomonas, Staphylococcus, Salmonella, and Enterococcus, were inhibited by the supernatant fluid. Bacteriocin is an antimicrobial peptide which was isolated from Lactobacillus delbrueckii. A single four-step process was carried out to purify it to homogeneity. MALDI-TOF investigation was performed with the crude supernatant acquired from an initial stationary phase culture in the MRS broth. Bacteriocin was proteolytic enzyme sensitive, heat resistant, functional over a broad pH range, and drifted as a peptide of 4.5–5.0. kDain the SDS–PAGE. During exponential growth, the bacteriocin was formed continuously. It was bactericidal to sensitive cells, and cell lysis did not achieve the bactericidal impact. The bacteriocin's amino acid composition was measured, and no modified amino acids were discovered among the residues found.