Various inorganic and organic nitrogen sources were used to compare their effects on the lipogenesis and the activities of lipogenic enzymes (providing acetyl-CoA and donating NADPH) in γ-linolenic acid-producing fungus Cunninghamella echinulata. Lipid accumulation was enhanced by organic nitrogen, among them the presence of corn-steep led to almost 40% oil in the biomass. While organic nitrogen increased activities of acetyl-CoA carboxylase (ACC) and malic enzyme (ME), ATP:citrate lyase (ACL) was rapidly enhanced by ammonium ion. The use of NaNO₃ resulted in high activities of glucose 6-phosphate dehydrogenase (GPD) and 6-phosphogluconate dehydrogenase (PGD). NADP-isocitrate dehydrogenase (NADP-ICD) was more active when the fungus utilized all inorganic N-compounds. The rise of nitrogen concentration in medium was accompanied with reduced lipid accumulation and a fall of ACL, ACC, and ME. In contrast, N-sufficient conditions favored biomass growth and elevated activities of GPD and PGD. Kinetic experiments also suggest that a significant portion of the required acetyl-CoA was being provided via ACL and ACC, and ME (probably coupled with GPD) channeled the NADPH into the fatty acid biosynthesis. The contribution of the lipogenic enzymes to metabolic pathways other than lipogenesis is also discussed.