Chlorella, among microalgae, has possibly been the most used for feeding, as well as the most researched in heterotrophic growing lately due to the highest lipid yield. The objective of this work was to study the effect of carbon and nitrogen concentration for lipid productivity and biomass production of microalga Chlorella minutissima, cultivated in absence of light. The complete factorial design used was type 22 with three repetitions at the central point, having as glucose concentration factor (1, 5 e 9 g.L-1) and as sodium nitrate (NaNO3) (0.2; 0.75 and 1.3 g.L-1), and total lipid and biomass concentration as responses, with a total number of 7 experiments. The cultivation carried out with 9 g.L-1 of glucose and 0.2 g.L-1 of NaNO3 was the one that presented better lipid concentration (29.38% w/w), better cell growth (0.89 g.L-1) and a lipid profile comprising saturated, monounsaturated and polyunsaturated fatty acids in 26.6% w/w, 62.4% w/w and 11% w/w concentrations, respectively. © All Rights Reserved.