Possible regulatory roles of ATP:citrate lyase, malic enzyme, and AMP deaminase in lipid accumulation by Rhodosporidium toruloides CBS 14

The properties of ATP:citrate lyase, malic enzyme, and AMP deaminase have been investigated in Rhodosporidium toruloides CBS 14. ATP:citrate lyase had a molecular size of 480 000 daltons and apparent K m for citrate and ATP of 0.19 mM and 0.15 mM, respectively. The enzyme was inhibited by ADP, glucose 6-phosphate, palmitoyl-CoA, and oleoyl-CoA. [Formula: see text] ions showed a 95% stimulation of activity at nonsaturating concentrations (0.1 mM) of citrate. Malic enzyme had a molecular size of 205 000 daltons and an apparent K m for malate of 0.7 mM. The enzyme was only weakly inhibited by citrate, pyruvate, oxaloacetate, and ATP but no metabolite was found which exerted a significant regulatory control over the enzyme. However this enzyme could be used as the principal, if not sole, source of NADPH needed for fatty acid biosynthesis. The role of this enzyme and the central position of malate as a key metabolite in determining how lipid accumulation could be initiated and then sustained is discussed. AMP deaminase was detected in low activities but was fourfold higher in nitrogen-limited cells. The possible role of this enzyme in degrading AMP, regulating cellular energy charge, and supplementing [Formula: see text] pools in this yeast is also discussed.