Microalgae, historically relevant to aquaculture sector, can effectively contribute to the sustainability in several production chain. The physical and chemical environmental factors play a direct or indirect role in shaping the composition of their intracellular compounds of these organisms. Among these compounds, fatty acids are particularly noteworthy, with various species harboring an extensive array of them. Notably, the marine dinoflagellate Prorocentrum micans excels in the production of essential polyunsaturated fatty acids (PUFA), including DHA and EPA. The objective was to determine which factors contribute to the accumulation of the PUFA in a P. micans strain. A factorial design of 3x3 composed by the following factors: illumination (5, 12.5, 20 µmol photons m-2 s-1), nitrate dose (0, 75, 150 mg ml-1) and inoculum volume (50, 125, 200 ml) was examined to determine the better growth conditions for P. micans. Furthermore, lipid profile and the fatty acids profile were analyzed at the end of the cultivation. The results determined that, maximum averages of DHA (20%) and EPA (1%), in the biomass, were registered in cultures without nitrate, low irradiance (5 µmol photons m-2 s-1) and high volume of inoculum (200 ml). The maximum cell concentration recorded was 3.1×104 cells ml-1, at the end of the culture. So, it can be concluded that the evaluated strain is easily adapted to the culture flow and, the culture conditions to which it was subjected allows its biomass to be considered an interesting alternative as lipid input in aquaculture activity.