Synthetic colorants have gradually been replaced by natural pigments, mainly in the food industry. Among the several natural sources of pigments, filamentous fungi have gained special attention. However, the eventual biosynthesis of mycotoxins is a limitation for the practical use of this kind of microorganisms as pigment producers. In this work, Penicillium brevicompactum was studied for the first time as a potential pigment producer using submerged fermentation. The effect of several experimental parameters (culture medium composition, agitation speed, type of carbon source, temperature, concentration of supplements and carbon source, natural light and initial pH) in the production of pigments was evaluated. Under the optimal conditions (culture medium I containing lactose (20g/L) and peptone/yeast extract (8/8g/L), 23°C, initial pH 7.0, 150rpm, and natural light), a mixture of pigments (yellow, orange, and red) was obtained. This mixture is mycotoxin-free (ochratoxin A and mycophenolic acid) and presented promising antioxidant activity (FRAP: 58.58±4.58µmol Fe(II)/g; DPPH IC50: 18.48±0.26µmol TE/g and ABTS IC50: 28.38±3.79µmol TE/g). Furthermore, these pigments were proved to be very stable for a wide range of pH and temperatures. Overall, it was demonstrated that P. brevicompactum can be a safe source of natural pigments with interesting properties for the food industry., This work was financially supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit. Luís Abrunhosa acknowledges FCT for the assistant research contract CEECIND/00728/2017, obtained under CEEC Individual 2017., info:eu-repo/semantics/publishedVersion