AbstractMicroorganisms are able to biosynthesize hydrocarbons (HCs), thus, they could become a new matrix for fuels and chemicals. In this work, Tetradesmus obliquus cultivated for 15 days in 12‐m3 photobioreactors produced dry biomass for hot oil extraction by rotary evaporation. Pure hexane and a mixture of hexane and ethanol solvents produced crude oil, followed by fractional distillation to yield non‐esterifiables. Gas chromatography‐mass spectrometry characterized the products obtained with hexane extraction as HCs from C11 to C22, and with hexane + ethanol from C13 to C23, predominantly alkanes that led to a high cetane number. The green diesel lower heating value was nearly the same as that of low sulfur fossil diesel, however, economic and logistic aspects need to be addressed aiming at fossil diesel replacement.