Enterococci have emerged as a global cause of nosocomial infection and are frequently associated with chronic ulcers, urinary tract infections, endocarditis and indwelling medical device-related infections. The robust biofilm formation and inherent multidrug resistance in Enterococci have uplifted it as a challenging nosocomial pathogen. Biofilms are cell populations irreversibly attached to surfaces and encased within a self-produced hydrated matrix of exopolymeric substances, proteins, polysaccharides, extracellular DNA (eDNA) and water channels. Biofilm confers high antibiotic tolerance ability and resistance to phagocytosis and hence play a prime role in critical colonization, establishment and chronicity of infections. This renders Enterococci to be recalcitrant towards the current treatment strategies. Owing to its egressing clinical significance, the pace of enterococcal biofilm research has accelerated in the past few years for providing a better understanding of intricate underlying mechanisms of biofilm formation.