Biosurfactants are naturally occurring amphiphiles that are being actively pursued as alternatives to synthetic surfactants in cleaning, personal care, and cosmetic products. On the basis of their ability to mobilize and disperse hydrocarbons, biosurfactants are also involved in the bioremediation of oil spills. Rhamnolipids are low molecular weight glycolipid biosurfactants that consist of a mono- or di-rhamnose head group and a hydrocarbon fatty acid chain. We examine here the micellization of purified mono-rhamnolipids and di-rhamnolipids in aqueous solutions and their adsorption on model solid surfaces. Rhamnolipid micellization in water is endothermic; the CMC (critical micellization concentration) of di-rhamnolipid is lower than that of mono-rhamnolipid, and both CMCs decrease upon NaCl addition. Rhamnolipid adsorption on gold surface is mostly reversible and the adsorbed layer is rigid. A better understanding of biosurfactant self-assembly and adsorption properties is important for their utilization in consumer products and environmental applications.