The antibacterial activities of the microbial pigment of the actinomycetes were carried out against the bacteria such as Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Escherichia coli, Salmonella typhi. The E. coli (8 mm) more susceptible than other microbial pathogens. Whereas remaining species showed negative result. The antifungal activity of the microbial pigment of the actinomycetes were carried out against the four different fungi such as Candida albicans, Aspergillus niger, Trichoderma viride, Rhizopus microporous. The actinomycetes pigment were failed to inhibit the pathogenic fungi. There is no antifungal activity against the all tested pathogenic fungi. Fourier Transform Infra-Red (FTIR) spectrum revealed the NH and NOH stretching for the actinomycetes pigment. In vitro cytotoxic assay on Human Cervical Adenocarcinoma (HeLa) cell line. The actinomycetes pigment prodigiosin, tripyrrole ring, was found to exhibit antiproliferative property against the (HeLa cell line. The pigment was found to be effective against the HeLa cell line. The IC50 value of the pigment was 62.50 μg/ml against HeLa cell lines respectively. From the results obtained it has been concluded that the pigment isolated from Streptomycetes could be used an antiproliferative agent. However it cannot be used as an antibacterial or antifungal agent.