AimThis research aims to characterize the pigment produced by isolated fungi and to evaluate its anticancer activities. Methods and ResultsPigment-producing fungi was isolated and identified as Fusarium chlamydosporum. The pigment was extracted with chloroform, purified by preparative TLC and characterized by Fourier transmittance infrared, electron spray ionization mass spectroscopy, liquid chromatography mass spectroscopy and nuclear magnetic resonance (NMR) ((HNMR)-H-1, C-13 NMR) spectral analysis, which revealed the pigment to be long chain hydrocarbons with poly unsaturated groups' (m/z 702). Pigment stability varied with different pH, temperature and sunlight conditions. The pigment-induced cell death in human breast adenocarcinoma cells MCF-7 and showed no significant toxicity in CHOK 1 cells. Lipid peroxidation assay revealed that treatment with pigment was able to reduce the lipid peroxidation caused by H2O2 in MCF-7 cells. ConclusionsThe F. chlamydosporum pigment is a compound of long-chain hydrocarbons with poly unsaturated groups, possessing selective cytotoxicity in MCF-7 cancer cell lines. Significance and Impact of the StudyThe pigment can be used as a colouring agent in cosmetics. Its anticancer potential can be used in production of therapeutics in increasing demand cancer research.