1887

Abstract

The distribution of carbon from glucose and glutamate was studied using anaerobically grown . The yeast was grown on glucose (20 g I-) as the carbon/energy source and glutamic acid (3.5 g I) as additional carbon and sole nitrogen source. The products formed were identified using labelled [U-C]glucose or [U-C]glutamic acid. A seldom-reported metabolite in 2–hydroxyglutarate, was found in significant amounts. It is suggested that 2-hydroxyglutarate is formed from the reduction of 2-oxoglutarate in a reaction catalysed by a dehydrogenase. Succinate, 2-oxoglutarate and 2-hydroxyglutarate were found to be derived exclusively from glutamate. Based on radioactivity measurements, 55%, 17% and 14% of the labelled glutamate was converted to 2-oxoglutarate, succinate and 2-hydroxyglutarate, respectively, and 55%, 9% and 3% of the labelled glucose was converted to ethanol, glycerol and pyruvate, respectively. No labelled glucose was converted to 2-oxoglutarate, succinate or 2-hydroxyglutarate. Furthermore, very little of the evolved CO was derived from glutamate. Separation of the amino acids from biomass by paper chromatography revealed that the glutamate family of amino acids (glutamic acid, glutamine, proline, arginine and lysine) originated almost exclusively from the carbon skeleton of glutamic acid. It can be concluded that the carbon flow follows two separate paths, and that the only major reactions utilized in the tricarboxylic acid (TCA) cycle are those reactions involved in the conversion of 2-oxoglutarate to succinate.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-144-6-1683
1998-06-01
2021-05-13
Loading full text...

Full text loading...

/deliver/fulltext/micro/144/6/mic-144-6-1683.html?itemId=/content/journal/micro/10.1099/00221287-144-6-1683&mimeType=html&fmt=ahah

References

  1. Achouri, Y., Rider, M. H., van Schaftingen, E., Robbi, M. (1997); Cloning, sequencing and expression of rat liver 3- phosphoglycerate dehydrogenase.. Biochem J 323,:365–370
    [Google Scholar]
  2. Albers, E., Larsson, C., Lid£n, G., Niklasson, C., Gustafsson, L. (1996); Influence of the nitrogen source on Saccharomyces cerevisiae anaerobic growth and product formation.. Appl Environ Microbiol 62,:3187–3195
    [Google Scholar]
  3. Avendano, A., Deluna, A., Olivera, H., Valenzuela, L., Gonzalez, A. (1997); GDH3 encodes a glutamate dehydrogenase isozyme, a previously unrecognized route for glutamate biosynthesis in Saccharomyces cerevisiae.. J Bacteriol 179,:5594–5597
    [Google Scholar]
  4. Bruinenberg, P. M., Dijken, J. P.v., Scheffers, W. A. (1983); A theoretical analysis of NADPH production and consumption in yeasts.. J Gen Microbiol 129,:953–964
    [Google Scholar]
  5. Buckel, W., Miller, S. L. (1987); Equilibrium constants of several reactions involved in the fermentation of glutamate.. Eur J Biochem 164,:565–569
    [Google Scholar]
  6. Christensen, L. H., Schulze, U., Nielsen, J., Villadsen, J.(199S). Acoustic off-gas analyser for bioreactors: precision, accuracy and dynamics of detection.. Chem Eng Sci 50,:2601–2610
    [Google Scholar]
  7. Cooper, T. G. (1982) Nitrogen metabolism in Saccharomyces cerevisiae. In The Molecular Biology of the Yeast Saccharomyces cerevisiae.. In Edited by Strathern, J. N., Jones, E. W., Broach, J. R. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory,;39–99
  8. Coote, N., Kirsop, B. H. (1974); The content of some organic acids in beer and other related fermented media.. J Inst Brew 80,:474–483
    [Google Scholar]
  9. DOrfel, H. (1959) Proteine und ihre Bausteine I. Aminosauren.. In Edited by Linskens, H. F. Papierchromatographie in der Botanik. Berlin:: Springer,;148–179
    [Google Scholar]
  10. Heerde, E., Radler, F. (1978); Metabolism of the anaerobic formation of succinic acid by Saccharomyces cerevisiae.. Arch Microbiol 117,:269–276
    [Google Scholar]
  11. Jansen, G. A., Wanders, R. J. A. (1993); L-2-Hydroxyglutarate dehydrogenase: identification of a novel enzyme activity in rat and human liver. Implications for L-2-hydroxyglutaric acidemia.. Biochim Biophys Acta 1225,:53–56
    [Google Scholar]
  12. Jones, E., Fink, G. (1982) Regulation of amino acid and nucleotide biosynthesis in yeast.. In Edited by Strathern, J. N., Jones, E. W., Broach, J. R. The Molecular Biology of the Yeast Saccharomyces cerevisiae. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;181–299
    [Google Scholar]
  13. Lewis, M. J., Rainbow, C. (1963); Transamination and the liberation of 2-oxoglutarate by yeast.. J Inst Brew 69,:39–35
    [Google Scholar]
  14. Miller, S. M., Magasanik, B. (1990); Role of NAD-linked glutamate dehydrogenase in nitrogen metabolism in Saccharomyces cerevisiae.. J Bacteriol 172,:4927–4935
    [Google Scholar]
  15. Nissen, T. L., Schulze, U., Nielsen, J., Villadsen, J. (1997); Flux distribution in anaerobic, glucose-limited continuous cultures of Saccharomyces cerevisiae.. Microbiology 143,:203–218
    [Google Scholar]
  16. Otawara, S., Ohshima, T., Esaki, N., Soda, K. (1984); Purification and properties of a-hydroxyglutarate dehydrogenase of Pepto- coccus aerogenes.. Agric Biol Chem 48,:1713–1719
    [Google Scholar]
  17. Perkins, M., Haslam, J. M., Linnane, A. W. (1973); Biogenesis of mitochondria. The effects of physiological and genetic manipulation of Saccharomyces cereviasiae on the mitochondrial transport systems for tricarboxylate-cycle anions.. Biochem J 134,:923–934
    [Google Scholar]
  18. Radler, F. (1986); Microbial biochemistry.. Experientia 42,:884–893
    [Google Scholar]
  19. Rankine, B. C. (1968); Formation of a-ketoglutaric acid by wine yeasts and its oenological significance.. J Sci Food Agric 19,:624–627
    [Google Scholar]
  20. Roels, J. A. (1983) Energetics and Kinetics in Biotechnology. Amsterdam:: Elsevier.;
    [Google Scholar]
  21. ter Schure, E. G., Sillj6, H. H. W., Raeven, L. J. R. M., Boonstra, J., Verkleij, A. J., Verrips, C. T. (1995); Nitrogen-regulated transcription and enzyme activities in continuous cultures of Saccharomyces cerevisiae.. Microbiology 141,:1101–1108
    [Google Scholar]
  22. Suzuki, T., Uozumi, T., Beppu, T. (1985); Purification and characterization of a-hydroxyglutarate dehydrogenase from Alcaligenes sp.. Agric Biol Chem 49,:2939–2947
    [Google Scholar]
  23. Wales, D. S., Cartledge, T. G., Lloyd, D. (1980); Effects of glucose repression and anaerobiosis on the activities and subcellular distribution of tricarboxylic acid cycle and associated enzymes in Saccharomyces carlsbergensis.. J Gen Microbiol 116,:93–98
    [Google Scholar]
  24. Woodward, J. R., Ciriilo, V. P. (1977); Amino acid transport and metabolism in nitrogen-starved cells of Saccharomyces cerevisiae.. J Bacteriol 130,:714–723
    [Google Scholar]
  25. Zhao, G. , Winkler, M. E. (1996); A novel a-ketoglutarate reductase activity of the serA-encoded 3-phosphoglycerate dehydrogenase of Escherichia coli K-12 and its possible implications for human 2-hydroxyglutaric aciduria.. J Bacteriol 178,:232–239
    [Google Scholar]
  26. Zhao, W.-N., McAlister-Henn, L. (1996); Expression and gene disruption analysis of the isocitrate dehydrogenase family in yeast.. Biochemistry 35,:7873–7878
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-144-6-1683
Loading
/content/journal/micro/10.1099/00221287-144-6-1683
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error