Abstract: The present study was an endeavour to select a probiotic bacteria inhibitory to human pathogenic bacteria. Characterization of bacteria by methods based on phenotypic, biochemical and molecular characters were followed herein and bacteriocin activity was a scope of the present study. Out of 20 isolates of lactic acid bacteria isolated from urine of healthy people, one isolate only: the NM2 strain inhibited other bacterial isolates isolated from patients with urinogenital infections and other bacterial pathogens of our strain collection. Based on biochemical, phenotypic characteristics and sequences of 16S rRNA gene the NM2 strain was identified as belonging to Enterococcus faecium (E. faecium NM2) and its two sensitive strains TW5; TCH4 were identified as Enterococcus faecalis (E. faecalis TW5); Burkholderia cepacia (B. cepacia TCH4). The inhibitory activity of cell free supernatant (CFS) of the NM2 strain was lost by proteolytic enzymes, heat resistant and was not affected by organic solvents, lipase and amylase. Consequently the active substance in CFS was proved to be of proteinaceous nature and was, therefore, characterized as a bacteriocin. This bacteriocin was active at acidic pH levels (pH 2.0-6.5) and its activity was lost at neutral and alkaline pH values; indicating on wider application of this bacteriocin.