Soil washing by surfactants is a technology to enhance mobilization and subsequent degradation of oil pollutants by reducing the surface tension of pollutants which is combined with soil. In this study, biosurfactant, rhamnolipid was produced from Pseudomonas aemginosa ATCC 9027 which had an excellent biodegradable activity in soil without causing secondary pollution. Effects of chemical surfactants on the removal of diesel from diesel-contaminated soil were compared to those of biosurfactants including rhamnolipid. Diesel removal efficiency by rhamnolipid extracted from P. aeruginosa culture broth was over 95% in both batch and column washing test in 5,000ppm diesel-contaminated soil with 1% surfactants after washing for 24 hours. On the contrary, the results of chemical surfactants were below 50∼80%, The chemical surfactants with HLB value(8∼15) showed more then 75% efficiency of diesel removal. But, when the HLB values were below 8 or over 15. their efficiency were observed as less then 60% of diesel removal. Rhamnolipid, biologically produced surfactants, may also be promising agent for enhancing diesel removal from contaminated soil.