Biofilm formation is closely related to the pathogenicity of E. faecalis in persistent root canal infections. Limited information is available about the ability and characteristics of E. faecalis biofilm-forming in the persistent pathogenicity of root canal infections. Based on these problems, the purpose of this paper is to improve the molecular understanding of E. faecalis on virulence factors associated with biofilm formation against persistent root canal infections to support laboratory diagnosis and therapy of oral E. faecalis. In conclusion, the release of cytokines triggers the dlt gene of LTA to improving: homeostasis, autolytic activity, and bacterial envelope properties. The role of cytolysin activated by the cylLL and cylLS genes improving the survival ability of E. Faecalis. Hyaluronidase will facilitate other bacteria to migrate from the root canal to periapical lesions resulting in the triggering of toxin production, which will increase damage and inflammation in the host. Enterococcus faecalis, through dentine matrix formation, hydrolyze collagen and induce apatite deposition in developing biofilms. Besides, these bacteria can also increase their tolerance to antimicrobials by blocking the inflammatory response's acid reaction. Alkaline conditions will neutralize the lactic acid secreted by osteoclasts to absorb hard tissue