Antibiotic resistance has emerged and persisted as one of the major threats to public health and the sustainable development of healthcare systems worldwide in recent years. The primary reason why antibiotic resistance is regarded as a global medical crisis lies in the development of new antibiotics is not keeping pace with the emergence of antibiotic-resistant strains. Currently, one of the proposed solutions to this crisis, as suggested by scientists, is the investigation of alternative antimicrobial agents that exhibit high efficacy while also possessing characteristics that limit the emergence of resistance in target pathogenic strains. Among such alternatives, endolysins, a class of enzymes derived from bacteriophages, have garnered considerable research interest. These enzymes are utilized by bacteriophages to degrade the peptidoglycan layer of bacterial cell walls during the infection process. The ability of endolysins to lyse bacterial cells when applied externally, combined with their high specificity at the genus or species level and a low propensity for inducing resistance, renders them promising candidates as substitutes for conventional antibiotics. This review aims to outline the structural features and mechanisms of action of endolysins, and to discuss in depth their characteristics and potential applications across various fields.