Open AccessA bacteriocin-producing strain isolated from Ethiopian honey was identified as Enterococcus faecium. The incidence of multidrug resistance has become a leading challenge in the treatment of deadly infectious diseases specifically of bacterial origin. In order to reduce the risk and spread of bacterial antibiotic resistance, various new drugs have been searched and introduced in the biomedical science. In this reference, bacteriocin could have been an important alternative for the control of certain infectious agents. The present study was carried out to check the effect of bacteriocin produced from Enterococcus faecium on multidrug resistant bacterial isolates. The isolates of Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, were isolated from different clinical specimens and were screened for multidrug resistance by the standard Kirby-Bauer disk diffusion method. Bacteriocin was produced from Enterococcus faecium during growth in MRS broth at 37°C and was checked for antibacterial activity against above mentioned resistant bacterial isolates by agar well diffusion method. All the clinical isolates were confirmed to be multi drug resistant. The bacteriocin proved to be active against a narrow range of gram positive bacteria i.e. isolates of Staphylococcus aureus and it showed reduced activity against; isolates of Pseudomonas aeruginosa and Escherichia coli.