Leishmaniasis is caused by protozoa of genus Leishmania, nowadays it affects millions of peoples worldwidly. Parasite proteases are described as essential in the host-parasite interaction. Extracellular proteases are considered important on immune system evasion and dissemination of protozoa on host. Our group manages to characterize extracellular proteases of L. amazonensis and L. chagasi. Considering the problems previously demonstrated by using bovine fetal serum (SFB) in culture, which carries many proteases and others interferent proteins, 10¹⁰ parasites were maintained in Schneider's Insect medium supplemented with 1% glucose for 12 hours at 26° C (SFB free) to evaluate proteases present in culture supernatant.  Cell free supernatant from L. amazonensis (PH8) and L. chagasi (PP 75) were obtained by centrifugation (3000g, 10', 4°C). The cell free culture supernatants were precipitated with (NH₄)₂SO₄ and the pellets were ressuspended, dialyzed in 10 mM Tris-HCl pH 7.5 and called extracellular extracts. We have examined activities of cysteine-, metallo-, serine-, and aspartic-proteinase by hydrolysis of specific chromogenic substrates at the optimal pH for each class. Our results show all enzymes activities studied in extracellular extracts from L. amazonensis and L. chagasi. Zimography using gelatin as substrate for Leishmania amazonensis showed apparent molecular weights of 92 KDa , 72 KDa, 63 KDa, 52 KDa and to Leishmania chagasi were detected activity bands with 82 KDa, 67 KDa, 61 KDa, 57 KDa, 45 KDa. In hemoglobin substrate gel electrophoresis, a 92 kDa band of L. amazonensis was detected. These bands were optimally active at pH 5.5. All bands were inhibited by 1,10-Phenanthroline, demonstrate that metallo proteases are the most abundant in culture supernatant. Effect of divalent ions (Ca⁺² and Zn⁺²) was analyzed and the presence of zinc ion inhibited all bands. To improve the characterization of extracellular proteases, immunoblot analysis was performed using anti-GP63 (Leishmanolysin), anti-cysteine-proteinase (anti-catalytic domain of Leishmania mexicana CPB), anti-papain, anti-calpain (Drosophila melanogaster calpain). Positive reactions were obtained against anti-GP63 (demonstrating an extracellular release of this enzyme) and anti-calpain.  Ours results indicate the presence of all class of proteases studied in extracellular extracts and the next step is to evaluate the participation of these proteases in host infection.