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Non Tuberculous Mycobacteria (NTM) comprises of more than 170 different species. Their resistance and lack of new therapeutic agents poses the challenge to the effective treatment of diseases caused by NTM pathogens. Mycobacterium kansasii is one of the most frequently isolated NTM which cause pulmonary diseases in humans. The treatment of M. kansasii disease includes multiple antimicrobials for the period of more than 12 months. However, their adverse effects and uncertain effectiveness remains challenging. Lactic Acid Bacteria (LAB) is a diverse group of bacteria who inhabits in a wide range of environment such as dairy, vegetables, gastrointestinal tract of humans, etc., They produces an antimicrobial peptide called Bacteriocins which are active against closely related species as well as others. In this study, we have screened the partially purified bacteriocins (PPB) extracted from LAB of cow milk samples against M. kansasii (MTCC3058). PPB of LAB strain BLN34 showed more than 50% of reduction in the growth of M. kansasii by Colony Forming Unit (CFU) estimation method and exhibits less in vitro toxicity in Vero cell lines. Also 50% of zebrafish larvae were survived upto 144 hpf in the presence of BLN34 PPB. The LAB strain BLN34 was identified as Enterococcus italicus by 16s rRNA analysis.