In the present work, the production of rhamnolipid from residual soybean oil (RSO) from food frying facilities was studied using a strain of Pseudomonas aeruginosa of contaminated lagoon, isolated from a hydrocarbon contaminated soil. The optimization of RSO, ammonium nitrate, and brewery residual yeast concentrations was accomplished by a central composite experimental design and surface response analysis. The experiments were performed in 500-mL Erlenmeyer flasks containing 50 mL of mineral medium, at 170 rpm and 30 +/- 1 degrees C, for a 48-h fermentation period. Rhamnolipid production has been monitored by measurements of surface tension, rhamnose concentration, and emulsifying activity. The best-planned results, located on the central point, have corresponded to 22 g/L of IZSO, 5.625 g/L of NH4NO3, and 11.5 g/L of brewery yeast. At the maximum point the values for rhamnose and emulsifying index were 2.2 g/L, and 100%, respectively.