Some of the benefits of the biosurfactants is that it can be produced from agroindustrial co product and is biodegradable. The soy molasses is a product co generated during soybean processing that has high production and low commercial value. This study investigated the use of Pseudomonas aeruginosa to produce biosurfactants in a soy molasses-based fermentation medium. Preliminary tests were performed to determine the best conditions of production. In these tests the bacterial cultures analyzed were: Pseudomonas aeruginosa ATCC 10145. After determining the best conditions it was developed a Central Composite Design (CCD) with two variables and three replicates at the central point to optimize the production of biosurfactant. The concentration of soy molasses has values between 29.3 and 170.7 g/L and the initial concentration of microorganism varies between 0.2 and 5.8 g/L. In response the following values were obtained at 48 hours of fermentation: emulsification index of 97.4% and concentration of biosurfactant of 11.7 g/L.