Tuberculosis is the principal cause of death from infection in the world. The resurgence of tuberculosis and the increase in mycobacterial infections, as well as multidrug-resistance of mycobacteria to available antibiotics, has incentivized research on new antimycobacterial agents. Therefore, research based on water and soil samples from the Moroccan biotopes, has led to the isolation of a bacterial strain capable of inhibiting mycobacterial growth (Mycobacterium smegmatis and Mycobacterium aurum A+). The effect was due to an active substance secreted into the culture medium. Sequencing of the 16S rRNA gene identified the strain as belonging to the species Staphylococcus warneri. The active substance precipitated using ammonium sulfate, maintained its inhibitory properties, which were lost when treated with proteinase K. These results indicated that the active substance was protein. Study of the activity of the metabolite revealed its effect on M. smegmatis cell wall, facilitating genomic DNA extraction.Keywords: Tuberculosis, mycobacteria, anti-mycobacterial agents, Staphylococcus warneri, DNA extraction.African Journal of Biotechnology Vol. 12(42), pp. 611