Lipopeptides are key elements in the formation of monolayer and bilayer phospholipid membranes, by which the dynamic strength, balance of acting forces, interaction modes, and arrangement patterns are maintained within phospholipid and epithelial cells. This review focuses on the production, structural characterization, and application of lipopeptides produced by various species of bacteria, which are grouped into three classes: surfactin, iturin, and fengycin. These compounds have tremendous potential in biomedicine. A cyclic lipopeptide correlated with peptides with heuristic structures of a functional spectrum, such as surfactin, the cyclic heptapeptide iturin, and the antifungal lipopeptide fengycin. Structural elucidation will be performed via advanced high-performance liquid chromatography, mass spectrometry, nuclear magnetic resonance, and Fourier transform infrared spectroscopy techniques to elucidate the peptide and fatty acid compositions of the lipopeptides in detail. This review has also considered the high-throughput screening methods thus far applied in identifying microorganisms that produce lipopeptides and the role of genetic manipulation in improving the productivity of lipopeptides. The elucidation of their structure and complex composition via chromatographic and spectroscopic techniques highlights the complexity of lipopeptide analysis. These biomolecules find applications as antibacterial, antiviral, and antifungal therapies in drug delivery systems and as bioactive agents in various industrial processes. Owing to their advantages in the encapsulation and transportation of bioactive substances, their prospective application in medicine and pharmaceuticals has increased.
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