Biosurfactants are chemical molecules produced by the microorganisms with potential for application in various industrial and environmental sectors. The production parameters and the physicochemical properties of a biosurfactant synthesized by Bacillus pumilus using different concentrations of vinasse and waste frying oil as alternative carbon sources were analyzed. The microorganism was able to grow and produce a biosurfactant using both the residues. The surface tension was reduced up to 45 mN/m and the maximum production of crude biosurfactant was 27.7 and 5.7 g/l for vinasse and waste frying oil, respectively, in concentration of 5%. The critical micelle concentration (CMC) results of 1.5 and 0.2 g/l showed the efficiency of the biosurfactant produced on both the substrates. The results showed that the alternative substrates could be used for the production of an efficient biosurfactant by B. pumilus. These properties have potential for industrial and environmental applications.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)