Biosurfactants have recently gained attention as "green" agents that can be used to enhance the remediation of heavy metals and some organic matter in contaminated soils. The overall objective of this paper was to investigate rhamnolipid, a microbial produced biosurfactant, and its ability to leach uranium present in contaminated soil from an abandoned mine site. Soil samples were collected from two locations in northern Arizona: Cameron (site of open pit mining) and Leupp (control-no mining). The approach taken was to first determine the total uranium content in each soil using a hydrofluoric acid digestion, then comparing the amount of metal removed by rhamnolipid to other chelating agents EDTA and citric acid, and finally determining the amount of soluble metal in the soil matrix using a sequential extraction. Results suggested a complex system for metal removal from soil utilizing rhamnolipid. It was determined that rhamnolipid at a concentration of 150 μM was as effective as EDTA but not as effective as citric acid for the removal of soluble uranium. However, the rhamnolipid was only slightly better at removing uranium from the mining soil compared to a purified water control. Overall, this study demonstrated that rhamnolipid ability to remove uranium from contaminated soil is comparable to EDTA and to a lesser extent citric acid, but, for the soils investigated, it is not significantly better than a simple water wash.