Abstract: The aim of the study was to assess the anti-tuberculosis potential of the Streptomyces species isolated from unexplored habitats. Six actinomycetes strains were screened for their anti-mycobacterial activity. The crude bioactive compound was prepared and extracted with resin Dianoin HP20. Actinomycete strains EWC 7(2), MN 2(6) and MN 9(V) showed prominent activity, and were further evaluated with BACTEC assay. The anti-mycobacterial activity was observed by detection of inhibition of growth in terms of growth index (GI value). M. tuberculosis H37 Rv and M. tuberculosis H37 Ra were found sensitive towards metabolites of Streptomyces sp. EWC 7(2), and Streptomyces sp. MN 2(6) as observed in the form of decreasing GI values. The findings of present study are promising however, further studies are required to purify and elucidate the structure of metabolites. These isolates may prove to be an important step in development of drug for treating mycobacterial infections.