Abstract
Surfactin, a group of cyclic lipopeptides produced by Bacillus subtilis, possesses surfactant properties and is a promising natural and biologically active compound. In this study, we present a comprehensive characterization of surfactin, including its production, chromatographic separation into pure homologues (C(12), C(13), C(14), C(15)), and investigation of their physicochemical properties. We determined adsorption isotherms and interpreted them using the Gibbs adsorption equation, revealing that the C(15) homologue exhibited the strongest surface tension reduction (27.5 mN/m), while surface activity decreased with decreasing carbon chain length (32.2 mN/m for C(12)). Critical micelle concentration (CMC) were also determined, showing a decrease in CMC values from 0.35 mM for C(12) to 0.08 mM for C(15). We employed dynamic light scattering (DLS), transmission electron microscopy (TEM), and density functional theory (DFT) calculations to estimate the size of micellar aggregates, which increased with longer carbon chains, ranging from 4.7 nm for C(12) to 5.7 nm for C(15). Furthermore, aggregation numbers were determined, revealing the number of molecules in a micelle. Contact angles and emulsification indexes (E(24)) were measured to assess the functional properties of the homologues, showing that wettability increased with chain length up to C(14), which is intriguing as C(14) is the most abundant homologue. Our findings highlight the relationship between the structure and properties of surfactin, providing valuable insights for understanding its biological significance and potential applications in various industries. Moreover, the methodology developed in this study can be readily applied to other cyclic lipopeptides, facilitating a better understanding of their structure-properties relationship.