Changes in free amino acid content and activities of amination and transamination enzymes in yeasts grown on different inorganic nitrogen sources, including hydroxylamine.
This study concerns inter- and intraspecific differences between yeasts at assimilation of different nitrogen sources. Alterations in the content of free amino acids in cells and media as well as in the related enzyme activities during growth were studied. The hydroxylamine (HA)-tolerant Endomycopsis lipolytica was examined and compared with the nitrate-reducing Cryptococcus albidus, and Saccharomyces cerevisiae, requiring fully reduced nitrogen for growth. Special attention was paid to alanine, aspartic acid, and glutamic acid, the amino acids closely related to the Krebs cycle keto acids. The amino acids were analyzed as their n-propyl N-acetyl esters by gas-liquid chromatography (GLC). The composition of the amino acid pool was similar for the three yeasts. Glutamic acid was predominant; in early log-phase cells of E. lipolytica contents of 200-234 micromol . g(-1) dry weight were found. A positive correlation between the specific growth rate and the size of the amino acid pool was observed. The assimilation of ammonia was mediated by glutamate dehydrogenase (GDH). The NADP-GDH was the dominating enzyme in all three yeasts showing the highest specific activity in Cr. albidus grown on nitrate (6980 nmol . (min(-1)).(mg protein(-1)). Glutamine synthetase (GS) displayed a high specific activity in S. cerevisiae, which also had a high amount of glutamine. The assimilation of HA did not differ greatly from the assimilation of ammonium in E. lipolytica. The existing differences could rather be explained as provoked by the concentration of available nitrogen.
D000410:Alanine Transaminase / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism*; D001203:Ascomycota / Q000378:metabolism*; D001219:Aspartate Aminotransferases / Q000378:metabolism; D003454:Cryptococcus / Q000378:metabolism*; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D006898:Hydroxylamines / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D004718:Saccharomycetales / Q000378:metabolism*; D013045:Species Specificity
D000596:Amino Acids; D006898:Hydroxylamines; D005969:Glutamate Dehydrogenase; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D005974:Glutamate-Ammonia Ligase