<p>Biosurfactants, amphiphilic molecules produced by microorganisms, are emerging as promising next-generation antimicrobials due to their diverse structures and mechanisms of action. This review explores the potential of biosurfactants in combating antimicrobial resistance (AMR) and their applications across various fields. We discuss the classification of biosurfactants, including glycolipids, lipopeptides, and phospholipids, highlighting their unique structural features and microbial origins. Key antimicrobial mechanisms are examined, focusing on membrane disruption, biofilm inhibition, quorum sensing interference, and immunomodulation. We detail the applications of biosurfactants in medicine, including their potential as therapeutic agents against multidrug-resistant pathogens and their use in drug delivery systems to enhance solubility and targeted delivery. Furthermore, we explore their agricultural applications as biocontrol agents and plant growth promoters, emphasizing their environmental compatibility. The review underscores the potential of biosurfactants to address the challenges posed by AMR and highlights future research directions, including genetic engineering, nanotechnology-based delivery, and clinical trials, to fully realize their therapeutic potential.</p> Graphical Abstract <p></p>