Biosurfactant produced from P. aeruginosa RB 28 was extracted, purified and characterized. Thin layer chromatography results showed that the extract contained two different compounds. The identification of the nature of the two compounds showed that they were glycolipids and rhamnose was the sugar moiety in these glycolipids. It was concluded that these compounds were rhamnolipids. The production of biosurfactant was started at late log phase and reached its maximal level (2.7 g L(-1)) at the stationary phase. Study of some rhamnolipid properties showed that sunflower oil, heptadecane and paraffin were efficiently emulsified and emulsions formed with vegetable oils (olive oil, corn oil and sunflower oil) were more stable than emulsions formed with hydrocarbons.