Abstract: Anti-Gardnerella vaginalis, Lactobacillus fermentum bacteriocin HV6b producing strain was isolated from vaginal swabs of healthy, fecund females. It was identified, purified and further characterized physicochemically. The relative molecular mass of bacteriocin was found to be less than 7 kDa. Maximum bacteriocin production was observed at 37°C, pH 6.5 after 16-18 h of incubation Proteinase K and pepsin strongly inhibited bacteriocin production. The bacteriocin has purified by absorption desorption method and characterized by reverse phase liquid chromatography. It exhibited high thermal stability and showed inhibitory activity over wide range of pH. The study revealed the possibility of using bacteriocin for bio-medical use and the L. fermentum strain as probiotic.