Abstract: Objective: To investigate the inhibitory effect of Enterococcus faecalis (EF162) on multidrug-resistant bacteria (MDRO) in vitro and its biological characteristics. Methods: Oxford cup agar diffusion method was used to determine the inhibitory effect of EF162 on eight strains of MDRO.Deere bacterial identification and antimicrobial susceptibility analysis system were used to determine the biochemical properties of EF162.Next-generation sequencing (NGS) was used to determine the whole genome sequence of EF162. Results: The strain was identified as Enterococcus faecalis and exhibited inhibitory activity against five strains of MDRO.NGS analysis revealed three potential bacteriocin-secreting gene clusters, which might encode cytolysins (ClyLl and ClyLs), Lanthipeptide and cochonodin Ⅰ. Conclusions: EF162 exhibits inhibitory effects against MDRO, possibly mediated through the production of Cytolysins (ClyLl and ClyLs), Lanthipeptide, and cochonodin Ⅰ.