Cholesterol (Chol) is a ubiquitous steroidal component of cell membrane and is known to modulate the packing of phospholipids within the bilayer. Thus, Chol has been frequently used in the formulation and study of artificial "model membranes" like vesicles and liposomes. In this work, we have developed a novel anionic surfactant by conjugating two biomolecules, cholesterol and gamma-aminobutyric acid via a urethane linkage. We have studied its physicochemical behavior in aqueous buffer. The surfactant has been shown to spontaneously form small unilamellar vesicles above a very low critical concentration in aqueous neutral buffer at room temperature. The vesicle phase was characterized by use of fluorescence probe, transmission electron microscopy and dynamic light scattering (DLS) techniques. The vesicle bilayer was found to be much less polar as well as more viscous compared to the bulk water. The vesicle stability with respect to change of temperature, pH, and ageing time was investigated by fluorescence probe and DLS techniques. The loading efficiency of the vesicles for the hydrophobic drug, curcumin, was determined and its release under physiological condition was studied. The in vitro cellular uptake of curcumin-loaded vesicles to human breast cancer cell line (MDA-MB-231) also was investigated. The MTT assay showed that the surfactant was non-cytotoxic up to a relatively high concentration. (C) 2017 Elsevier Inc. All rights reserved.