It is well known that structural factors affect hydration-induced amphiphile self-assembly through differences in molecular shape and the subsequent intrinsic curvature of the molecules. The effect of stereochemistry on the characteristics of self-assembled structures has been demonstrated experimentally with gemini surfactants and sphingomyelin. In the present study, the MMG-6 stereoisomers adopted more splayed and disordered tail configurations with higher APL. It has been proposed that the native-like lipid acid configuration allows MMG-6 hydrocarbon chains to take conformations with larger spatial volumes in comparison to those of MMG-1. 11 Both wide and small angle X-ray scattering studies have previously shown that MMG-6 forms an inverted hexagonal (H II ) phase at room temperature, whereas MMG-1 maintains planar structures at temperatures of up to 60°C. As the H II phase arises from the relatively small headgroup to tail ratio and subsequent spontaneous negative curvature induced for the monolayer, our results support the idea of a larger spatial volume occupied by the MMG-6 tails. Furthermore, previous Langmuir monolayer studies have indicated that a higher surface pressure is required for MMG-6 to transition from the liquid-expanded to the liquid-condensed phase compared to the case for MMG-1. 10 This requirement for a higher surface pressure was attributed to the more wedge-shaped molecular structure of MMG-6, which restricts the formation of similar favorable hydrophobic interactions observed for MMG-1. Our simulation results align with this perspective, as evidenced by consistent observations in the order parameters, splay angle, and g 3 clustering; all of these analyses indicate that the acyl chains of MMG-6 lipids display higher acyl chain entropies, which effectively increases the volume adopted by the acyl chains and possibly hinders acyl chain packing.