Tuberculosis and leprosy, caused by Mycobacterium tuberculosis and Mycobacterium leprae, respectively, are still very prevalent diseases in the world. However, the responsible mechanisms for the pathogenesis of these organisms are largely unknown. Microbial pathogens frequently utilize extracellular proteases as virulence factors, which often contribute significantly to pathology. These proteases participate in tissue destruction, inactivation of host defense molecules, activation of key regulatory proteins or peptides, nutrient acquisition and the processing of secreted signaling molecules that regulate gene expression. The analysis of both M. leprae and M. tuberculosis genomes revealed the presence of a large number and variety of putative protease genes, some encoding potential secreted proteases. Despite this, only a few mycobacterial proteases have been investigated. In a previous study, we have described the cloning of a putative secreted glycoprotease (Gcp) (ML0379). A Gcp homologous to ML0379 have been described in Pasteurella haemolytica A1 associated with bovine pneumonic pasteurellosis pathogenesis. Here we report the purification of the M. leprae recombinant Gcp. M. leprae Gcp was His-Tagged purified using a Ni-NTA column under denaturation conditions. The purified rGcp was used to obtain a specific polyclonal antibody in mouse. This antibody will be useful to monitor the expression of Gcp by M. leprae and M. tuberculosis both in vitro and during infection. We also cloned the M. tuberculosis Gcp homolog (Rv3419c) that shares 86% Identity with M. leprae Gcp. The M. tuberculosis Gcp was cloned into the vectors pVV16 and pVV16ap that allow expression both in Escherichia coli and mycobacteria. rRv3419c was constitutively expressed in Mycobacterium smegmatis mc²155. Currently the enzymatic activity of rGcp is under investigation.