Pigment from the endophyte Burkholderia sp. WYAT7 isolated from the medicinal plant Artemisia nilagirica (Clarke) Pamp. was extracted. The antibacterial efficacy of the crude pigment Y was assessed as a source of antibiotic against both Gram-positive and Gram-negative bacterial pathogens. The pigment Y exhibited a significant level of antibacterial activity against the tested pathogens Salmonella typhi (MTCC 733), Staphylococcus aureus (MTCC 1430), Pseudomonas aeruginosa (MTCC 2453), Klebsiella pneumoniae (MTCC 432), Escherichia coli (MTCC 1610), Salmonella paratyphi (3220), Bacillus subtilis (441) and Acinetobacter baumannii (12,889). The minimum inhibitory concentration of crude pigment extract Y for most of the bacterial pathogens tested was below or equal to 0.25 mu g/mL and the minimum bactericidal concentration was below or equal to 0.5 mu g/mL. In the cytotoxicity evaluation, crude pigment Y exhibited less toxicity toward normal cells lines (L929). Crude pigment extract Y also showed powerful anticancer activity toward melanoma cancer cells (A375). The IC50 value obtained was 68.08 mu g/mL. Acridine orange (AO) and ethidium bromide (EB) double staining of cells treated with the pigment helped in the morphological assessment of nuclear condensation, apoptotic bodies and live cells. The DNA fragmentation analysis and caspase-9 quantification in the pigment-treated A375 cells determined the apoptosis activity mediated by the crude pigment extract Y. The compounds in the crude pigment extract Y was identified by HR-LCMS analysis. Further studies on the active compounds can lead to a rise in new drugs for cancer treatment and also against rising antibacterial resistant pathogens.