Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) ; The use of biodiesel as an alternative to fossil fuels has been increasing in recent years. Since each ton of biodiesel is generated 100 kg of glycerol, there is some concern about the destination of this byproduct, as its disposition is economically expensive and its market value is low. This way, a solution to this problem is the use of glycerol as a source of carbon in the production of biosurfactants. Despite the numerous advantages of biosurfactants, such as low toxicity, biodegradability and high stability, these compounds are not widely used due to the high cost of production. The application of alternative carbon sources can contribute to reduce production costs and increase the commercial use of biosurfactants. The aim of this research was to analyze the potential of Biodiesel glycerol as carbon source in the production of rhamnolipids from Pseudomonas aeruginosa LBI 2AI. Five samples of glycerol, including treated and untreated, were evaluated. The experiments were conducted in flask and Bioreactor. The 1L Erlenmeyer flask containing 300 mL of culture medium were incubated in a rotary shaker for 120 hours at 37°C and 200 rpm. Cultures in 13L bioreactor with 4L of culture medium were carried out for 160 hours at 37°C. The agitation speed varied between 300-900rpm in order to maintain pO2 in 5%. To improve the biosurfactant concentration, fed-batch culture was performed witch up to three pulses of glycerol feeding, inceasing the concentration of the medium by up to 30% of initial. The results show that Pseudomonas aeruginosa LBI 2AI was able to produce rhamnolipídios from residual glycerol from Biodiesel production in the strategies used in this study. Bioreactor cultures enabled an increase in the concentration of the product formed, using the treated and untreated Glycerol residues. Promising concentrations of rhamnolipids using these carbon sources were achieved increasing the concentration. ; A utilização do biodiesel como alternativa em ...