The aims of this study were to identify bacteriocinogenic activity in 13 enterococci isolated from fecal samples of wild South American (Arctocephalus australis) and Subantarctic fur seals (Arctocephalus tropicalis); to determine the physicochemical characteristics and antimicrobial spectrum of antimicrobial compounds against Gram-positive and Gram-negative bacteria; and to evaluate the presence of bacteriocin structural genes by PCR. Out of 13 enterococci screened for antimicrobial activity, five enterococci showed activity against Listeria monocytogenes ATCC 35152, an important pathogen linked to food. Of these, only the E. mundtii strain J5 maintained the activity after the pH was adjusted (pH 6.5). The activity of antimicrobial compounds from the E. mundtii strain J5 (ACs-J5) was lost after proteolytic enzyme treatment; however, the activity was maintained after heat, pH (acidic and basic conditions) and chemical treatment. ACs-J5 showed narrow spectrum activity. Only the mundticin KS gene was detected in the J5 strain and no plasmid was present. In conclusion, the properties presented by ACs-J5 make it a valuable biopreservative in food industries in avoiding pathogenic microorganisms such as L. monocytogenes and it should be a good candidate for probiotic application.