In this work, the thermodynamics of molecular self-assembly for an anionic biosurfactant, sodium deoxycholate (NaDC) and their mixture with an oppositely charged surfactant, docecyltrimethylamonium bromide (DTAB) in aqueous solution was investigated by isothermal titration calorimeter (ITC), companied with turbidity and conductivity measurements. From the perspective of interaction energy, the results for single surfactant NaDC obtained from ITC provided a direct evidence for the stepwise association model to form the premicelles in the first step and the stable micelles in the second step. We obtained a series of thermodynamic parameters of premicellar and micellar formation, and thus discussed the thermodynamic mechanisms of molecular aggregation in these two processes. Furthermore, we investigated the thermodynamics of intermolecular interaction for the mixed systems composed of oppositely charged surfactants (DTAB/NaDC). The mixed critical micelle concentration (cmc mix ) and the transition concentration between two micellar morphologies (C M ) and their corresponding changes of enthalpy in the both processes were derived from ITC measurements in rich-NaDC and in rich-DTAB region, respectively. The results show that aggregation behaviors of DTAB/NaDC differ from single surfactant system or the normal mixed surfactants with charged "head-and-tail", due to the asymmetry in molecular structures caused by the rigid steroid skeleton of NaDC composed of a hydrophilic and a hydrophobic surfaces. As a result, the occurrence of various aggregation processes in mixed solution can be controlled by the total concentrations of two surfactants and their molar ratio. Conclusively the process of mixed micellization is driven by entropy, and the transition between two kinds of micellar morphologies is driven jointly by entropy and enthalpy. Therefore, from the perspective of interaction themodynamics, this work gave a deep insight in understanding the themodynamics of molecular selfassembly and their phase behavior for mixed systems with bile salts and the positively charged surfactant.