Tuberculosis is an infectious disease that causes fatality every year because of drugs resistance of the Mycobacterium complex; this is why the development of new antibiotics becomes an urgent need. To reach that target, a main approach was led; consisting on screening of active substances producing microorganism. In this study, we reported data on a strain that was isolated from different areas of Fez (Morocco), which present an antagonistic effect against Mycobacterium smegematis and possessed a large spectrum against bacteria of Gram positive and negative. The antimycobacterial compounds producer, ZI1, was identified as Aerococcus sp. on the basis of PCR amplification of 16S ribosomal RNA gene followed by sequencing (99 and 100% of homology, respectively by RS16 and FD1), and by using biochemical tests, the unknown bacterium ZI1 was so close to either the species Aerococcus viridans or Aerococcus urinaeequi, but it was distinguishable from the other five Aerococcus species. The antimycobacterial compounds were synthesised in the exponential growth phase of Aerococcus sp. and were fully affected following heat treatment and protolytic enzymes which indicated the proteinaceous nature of the active agents.