The biosurfactants have many advantages in comparison to the sinthetic surfactants, such as low toxicity, biodegradability, and chemical and structural diversity. The purpose of this work was to study the application of the method of the oil spreading to quantify the production of surfactants by different strains of Pseudomonas [P. putida, P. aeruginosa, P. fluorescens, and three isolates of Pseudomonas spp. (Iso 1, Iso 2 and Iso 3)], and also to verify the extracellular secretion of some enzymes (amylases, pectinases, nucleases, proteases and lipases) of the same microorganisms. The composition of the growth medium used in the oil spreading method was (g L^-1): 2.7 KH₂PO₄; 13.9 K₂HPO₄; 10.0 sacarose; 50.0 NaCl; 0.5  yeast extract; 1.0 NaNO₃; after the autoclavation of this, it was added the follow autoclaved aqueous solutions (10 mL L^-1) A (25.0 gL^-1 MgSO₄) and B [100.0 gL^-1 (NH₄)₂SO₄], and the filtered (0.22 mm) solution C (gL^-1: 0.5 EDTA; 3.0 MnSO₄.H₂O; 1.0 NaCl; 0.1 CaCl₂.2H₂O; 0.1 ZnSO₄.7H₂O; 0.1 FeSO₄.7H₂O; 0.01 AlK(SO₄)₂; 0.01 Na₂MoO₄.2H₂O; 0.01 H₃BO₃; 0.00001 CuSO₄.5H₂O). This is a modified composition of the medium described by Yossef et al.(2004).  Then, the optical density _600nm, and the number of cells mL^-1 were determined at intervals of 24 h, during 120 h. Aliqüotes of the filtered growth medium were also analysed by the mineral oil spreading method, after the construction of a standard courve using different concentrations (0-1000 mgL^-1) of a sinthetic surfactant (Duodecil Sulfate of Sodium - SDS) versus the diameter of the degradation of mineral oil. The maximum concentrations of the biosurfactant 120 h after the inoculation were 247 eq. mg of SDSL^-1 for Iso 1;  180 eq. mg of SDSL^-1 for Iso 2, 178 eq. mg of SDSL^-1 for P. putida; 170 eq. mg of SDSL^-1 for P. fluorescens; 166,2 eq. mg of SDS L^-1 for P. aeruginosa;  and 148,4 eq. mg of SDS L^-1 for Iso 3. The increase of the optical density was parallel to the surfactant production during the incubation time. Regarding the tests using different substrates in solid growth medium, 72 h after the incubation (30 ­+ 1 ºC, dark), all the strains produced the analyzed enzymatic activities. Then, the tested strains of Pseudomonas, mainly the Iso 1, showed potencial to be used in combination  with other microorganisms in assays of bioremediation of industrial effluents containing liposoluble compounds. Financial Support: Finep/CTHidro; BNB; Capes; CNPq; Fapeal; S.A. Usina Coruripe Açúcar e Álcool.