Rhamnolipids are biosurfactants substances produced by bacterial fermentation process of Pseudomonas aeruginosa, required for the remobilization of oil and used for fight against pollution. In the present study, 26 bacterial strains were isolated from soil and sewage of two different sites polluted by hydrocarbons, in which 3 selected strains producing Rhamnolipids belonging to the genus of Pseudomonas aeruginosa were purified, identified and used for further characterization. The secondary screening of the selected Pseudomonas aeruginosa revealed that one strain manifested a high performance for Rhamnolipids production. For the optimization of several parameters implicated in the biosurfactants production, five carbon sources such as olive oil, sunflower oil, glycerol, ethanol, corn germ and six nitrogen sources such as yeast extract, soybean meal, peptone, meat extract, urea, corn steep liquor has been investigated. The obtained results showed that the maximum Rhamnolipids production (2.7 g/l) was yielded by the using olive oil at a concentration of 3% as carbon source and 5 g/l of extract yeast as nitrogen source.
