<b>Background and Objective:</b> Preservation using antimicrobials has been observed to inhibit the growth of pathogenic bacteria in food. Nowadays many people choose food preservatives that are safe for health and natural. Bacteriocins as food preservatives are safe because antimicrobials from the antimicrobial peptide group include GRAS (Generally Recognized As Safe). Bacteriocin-producing LAB can be found in various fermented foods, one of which is "Dadih". Bacteriocins are expected to inhibit the growth of pathogenic bacteria so that they can be developed as an alternative to food preservatives. <b>Materials and Methods:</b> In this study, all experiments were performed with two replicates and the results were expressed as Mean±Standard Deviation (SD). <b>Results:</b> Screening results showed that the DK8 isolate had the highest antimicrobial activity. The DK8 isolate was identified molecularly using 16s RNA sequencing, showing that the DK8 isolate had the highest similarity to <i>Lactobacillus pentosus</i> strain 124-2. Bacteriocins from DK8 isolate and partially purified using ammonium sulfate precipitation at concentrations of 50, 60 and 70%. The addition of ammonium sulfate with a concentration of 50% showed the highest antimicrobial activity against <i>Salmonella</i> sp. (12.63 mm) and <i>Escherichia coli</i> (11.33 mm) while the highest antimicrobial activity against <i>Staphylococcus aureus</i> was the addition of 60% ammonium sulfate (8.13 mm). <b>Conclusion:</b> Lactic acid bacteria isolate was identified to have the highest similarity with <i>Lactobacillus pentosus</i> strain 124-2 and precipitation using 50% ammonium sulfate showed the highest antimicrobial activity.