DNA damaging activity of anticancer drugs produced by Actinomycetes is directly linked to the cell cycle arrest leading to apoptosis and other secondary responses.Major signaling pathways are involved for permeability of mitochondrial membranes and the binding of receptor proteins leading to cell death. Extracellular compounds from Actinomycetes cells expressed higher levels of the anti- tumour enzyme (290µg/ml/hr) which induces considerable damage to cancer cell lines. L-asparaginase activity reported is higher than those expressed by the microbes currently used for commercial production. A very low percentage of viability was noted for the cancer cells when compared to the normal controls. DNA breaks are observed in the tetrads of cancer cells when treated with the exudates but the normal cells have not been disturbed. Studies on the exudates have identified novel compounds which have proven anti- tumour property. The 16s rRNA sequencing confirmed the strains involved to be of Actinomycetes, including three novel strains. The DNA cleaving properties of the Actinomycete exudates have opened up possibilities to limit cell proliferation in Cancer cell lines.