Studies on the control and properties of ornithine transcarbamylase in germinated spores of Geotrichum candidum

The effects of arginine and urea on the levels of ornithinetranscarbamylase (OTC) were investigated in relation to thephysiological functions of this enzyme in Geotrichum candidum.OTC was repressed in germinated spores to half of its initiallevel when exogenous arginine exceeded 12 mM. The repressionof OTC could not be correlated with intracellular arginine concentration.The addition of urea at the final concentration of 0.035 M increasedthe specific activity of OTC by 5.5 and 2.5 fold as comparedto enzyme levels in arginine-repressed spores and control sporesrespectively. Simultaneous addition of urea and arginine duringgermination prevented either arginine repression or urea inductionof OTC. The enzyme was partially purified from germinated sporesand isolated as a single protein band after disc electrophoresis.Two distinct pH optima for the forward reaction (pH 8.8–9)and backward reaction (pH 7.8) were found. Km values for ornithineand carbamyl phosphate were 5 x 10^–3 M and 6.8 x 10^–4so respectively. The Km for citrulline in the catabolic directionwas 1 x 10^–2 M. Enzymes obtained from cell-free extractsof germinated spores could synthetize ATP from citrulline andADP under physiological conditions by coupling the phosphorolysisof citrulline with carbamate kinase activity. The initial rateof germination was stimulated in the presence of citrullineas the sole nitrogen source, as compared to arginine, glutamineor yeast extract. These observations suggest that citrullinemay be catabolized during germination by means of OTC ratherthan via the energy-consuming urea cycle. (Received June 26, 1971; )