-Biosurfactants have a wide range of applications in emulsions, separations, and solubilization because these chemicals reduce the surface tension and viscosity of solutions. This work studied rhamnolipid production using a batch bioreactor with a working volume of 1.5 liters, Pseudomonas aeruginosa ATCC (American Type Culture Collection) 10145 strain, and the greasy effluent from the slaughter of poultry and pigs as the substrate. The main goal of this research was to evaluate the level of aeration, agitation speed and inoculum concentration using a Central Composite Design (CCD). Experimental conditions were selected using the surface response technique obtained from the CCD, and the results were validated to test the reproducibility. The following operating conditions were selected: 1.2 vvm level of aeration, 600 rpm agitation speed, and 1.0 g/L biomass inoculum concentration. Under these conditions, the following results were obtained: the rhamnose production, surface tension and emulsifying index were 5.37 g/L, 25.6 dyne/cm and 100%, respectively.