Summary
Dialyzed cell-free preparations of baker's yeast fortified with magnesium and potassium ions, CoA and ATP incorporate 14C-labeled acetate in the presence of unlabeled α-ketobutyrate. This acetate-fixing reaction results in the formation of only one product that has been isolated by paper chromatography and is catalyzed by an enzyme which condenses in the absence of magnesium ions 1 mol of acetyl-CoA with 1 mol of α-ketobutyrate. The new condensing enzyme is very active in the crude extracts and has been separated by ammonium sulfate fractionation from other enzymes previously reported to occur in baker's yeast, which condense acetyl-CoA with the following α-ketoacids: glyoxylate, pyruvate, oxaloacetate, α-ketoisovalerate, and α-ketoglutarate.
Zusammenfassung
Dialysierte zellfreie Extrakte aus Bäckerhefe fixieren, mit Hilfe von Magnesium und Kaliumionen, Coenzyme A und ATP, 14C-markiertes Acetat in Gegenwart von unmarkiertem α-Ketobutyrat. Diese Acetat-Fixierungsreaktion führt zur Bildung eines einzigen Produktes, das durch Papierchromatographie isoliert worden ist, und wird von einem Enzym katalysiert, das, in Abwesenheit von Magnesiumionen, 1 Mol Acetyl-CoA mit 1 Mol α-Ketobutyrat kondensiert. Das neue kondensierende Enzym ist sehr aktiv in Rohextrakten und konnte durch Ammonsulfatfraktionierung von anderen schon beschriebenen Hefeenzymen, welche die Kondensation von Acetyl-CoA mit verschiedenen α-Ketosäuren, nämlich Glyoxyl-, Brenztrauben-, Oxalessig-, α-Ketoisovalerian- und α-Ketoglutarsäure, durchführen, getrennt werden.
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This investigation was supported by Grant No. AM 06848-02 from the National Institutes of Health, United States Public Health Service.
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Canovas, J.L., Ruiz-Amil, M. & Losada, M. Condensation of α-ketobutyrate and acetyl-CoA in baker's yeast. Archiv. Mikrobiol. 50, 164–170 (1965). https://doi.org/10.1007/BF00409126
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DOI: https://doi.org/10.1007/BF00409126