The problem of heavy metals contamination is a global issue and the challenge is to develop methods to remove heavy metals from soil and water. Recently, technologies using microorganisms and microbial products to remove metals have been successfully applied to waste water streams such as sewage sludge, and industrial effluents. Anionic bio-surfactants (Rhamnolipids) produced by the bacterium of Pseudomonas species has been effectively used as heavy-metal cleanup. This study investigates the feasibility of using bio-surfactants (Rhamnolipids) to remove or reduce heavy metals (Cd2+, Pb, Ni2+, Ba, Zn and Sr). Different concentrations (20, 40, 80 ppm) of Rhamnolipids were evaluated and compared for the efficiency of heavy metals removal. Results showed that, 80 ppm of bio-surfactants were required to reduce up to (53%, 62%, 56%, 28%, 20% and 7%) of Cd2+, Pb, Ni2+, Ba, Zn and Sr respectively. Rhamnolipid at concentration of 80 ppm could be successfully used as bio-surfactants to remove heavy metals from contaminated water.