A metabolomic study of the effect of glutamate dehydrogenase deletion on growth and morphogenesis.

There are two glutamate dehydrogenases in the pathogenic fungus . One is an NAD-dependent glutamate dehydrogenase () and the other is an NADPH-dependent glutamate dehydrogenase (). These two enzymes are part of the nitrogen and nicotinate/nicotinamide metabolic pathways, which have been identified in our previous studies as potentially playing an important role in morphogenesis. In this study, we created single gene knockout mutants of both dehydrogenases in order to investigate whether or not they affect the morphogenesis of . The genes were deleted and the phenotypes of the knockout mutants were studied by growth characterisation, metabolomics, isotope labelling experiments, and by quantifying cofactors under various hyphae-inducing conditions. We found that the mutant was unable to grow on either arginine or proline as a sole carbon and nitrogen source. While the / mutant could grow on these carbon and nitrogen sources, the strain was locked in the yeast morphology in proline-containing medium. We detected different concentrations of ATP, NAD, NADH, NAPD, NADPH, as well as 62 other metabolites, and 19 isotopically labelled metabolites between the mutant and the wild-type strains. These differences were associated with 44 known metabolic pathways. It appears that the disequilibrium of cofactors in the / mutant leads to characteristic proline degradation in the central carbon metabolism. The analysis of the / and the / mutants confirmed our hypothesis that redox potential and nitrogen metabolism are related to filament formation and identified these metabolic pathways as potential drug targets to inhibit morphogenesis.