The trade of healthy food and its demand for human welfare endorse the researchers to find novel and potent antimicrobial compounds for food preservation. The present investigation focuses on the purification and characterization of a novel bacteriocin produced by Acinetobacter movanagherensis AS isolated from goat rectum. Initially, 28 bacterial cultures having antimicrobial activity were isolated from animal organs like intestine and rectum of chick, goat, pig and sheep. Among them, a potent bacteria isolated from goat rectum was selected on the basis of its antimicrobial potential against Gram positive (Staphylococcus aureus and Listeria monocytogenes) and Gram negative bacteria (Salmonella typhi, Shigella boydii, Escherichia coli and Vibrio parahaemolyticus). Potent organism was identified as Acinetobacter movanagherensis AS with the help of biochemical and 16S rDNA sequencing. Tryptic soy broth supplemented with 1% yeast extract having pH 6.5 incubated at 37°C for 32 h were the optimum conditions for bacteriocin production. 70% ammonium sulphate precipitated and dialysed sample was purified using DEAE cellulose and sephadex G75 column chromatography. Bacteriocin was purified by 23.43 fold with 7.93% recovery and its molecular weight was found to be 12.11 kDa by SDS-PAGE analysis. Its biochemical characterization showed good stability at broad range of temperature (40-80ºC) as well as pH (3-9). Inhibition of antimicrobial activity was observed after treatment with trypsin, pepsin and protease which confirmed the proteinaceous nature of the purified bacteriocin. Bioinformatic analysis suggests that the purified bacteriocin under investigation may belong to colicin family. Considering the antimicrobial potential of purified bacteriocin coated on LDPE film, we assume that it can be potentially exploited for food preservation in food processing industries. Keywords: Bacteriocin; Acinetobacter movanagherensis; Column chromatography; LDPE Film.