During the biodiesel synthesis, there is the emergence of co-products dissolved at the reaction, thus, it is necessary that a separation process of these co-products occurs, generating an effluent that is considered polluting. This work aimed to characterize the biodiesel effluent, besides evaluating the alternative treatment of this medium using a biotechnological method. The parameters: pH, turbidity, conductivity, COD, BOD5, total solids, total dissolved solids, temperature and content of oils and greases. During the research was carried out an experimental design for the application of the Rhizopus oryzae URM 4692 lipase for the treatment of biodiesel effluent. In this process, pH, application concentration of gum arabic (7% w/v) and the enzymatic charge were used. It should be noted that the removal of the COD content of the effluent was evaluated in this statistical modeling. Experimentally, it was observed that the collected effluent presented high content of oils and greases, COD and organic load. It was possible to verify that all the variables were significant, but the concentration of lipase application was the most evidenced, since the tests that used the highest concentration of this variable obtained the lowest COD results, Reaching 207,783.6 mg.L-1 or 29.4% removal efficiency of this parameter.