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Pyruvate Oxidase

  • Robert B. Gennis
  • Lowell P. Hager

Abstract

Pyruvate oxidase (pyruvate: cytochrome b 1 oxidoreductase, EC 1.2.2.2) is a peripheral membrane enzyme isolated from Escherichia coli which catalyzes the oxidative decarboxylation of pyruvate to yield acetate plus CO2. This enzyme is water-soluble and is removed from the bacteria by sonic oscillation. Relatively large quantities of the oxidase can be purified to homogeneity, and it is reasonably stable. Thiamine pyrophosphate (TPP) and flavin adenine dinucleotide (FAD) are known cofactors. The reduced enzyme (FADH2) can be coupled to a crude membranous particulate fraction from E. coli containing an electron transport chain in which oxygen is the terminal electron acceptor. Thus, either CO2 release or O2 consumption may be used to assay for oxidase activity in this system. The reduced enzyme will also directly reduce the dye 2,6-dichloroindophenol (DCIP), or ferricyanide, providing convenient spectrophotometric assays.

Keywords

Biological Chemistry Flavin Adenine Dinucleotide Trypsin Activation Crude Lipid Flavin Adenine Dinucleotide 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Cunningham, C. C., and Hager, L. P., 1971a, Crystalline pyruvate oxidase from Escherichia coli: Activation by phospholipids, J. Biol. Chem. 246:1575–1582.PubMedGoogle Scholar
  2. Cunningham, C. C., and Hager, L. P., 1971b, Crystalline pyruvate oxidase from Escherichia coli: Phospholipid as an allosteric effector for the enzyme, J. Biol. Chem. 246:1583–1589.PubMedGoogle Scholar
  3. Gounaris, A. D., and Hager, L. P., 1961, A resolution of the Escherichia coli pyruvate dehydrogenase complex, J. Biol. Chem., 236:1013–1018.PubMedGoogle Scholar
  4. Hager, L. P., 1957, Trypsin activation of a ferricyanide-linked pyruvic acid oxidation, J. Biol. Chem. 229:251–263.PubMedGoogle Scholar
  5. Kaback, H. R., 1974, Transport studies in bacterial membrane vesicles, Science 186:882–892.PubMedCrossRefGoogle Scholar
  6. Kaczorowski, G., Shaw, L., Fuentes, M., and Walsh, C., 1975, Coupling of alanine racemase and D-alanine dehydrogenase to active transport of amino acids in E. coli B membrane vesicles, J. Biol. Chem. 250:2855–2865.PubMedGoogle Scholar
  7. Sweeney, E. W., 1961, M.S. Dissertation, University of Illinois.Google Scholar
  8. Williams, F. R., and Hager, L. P., 1966, Crystalline flavin pyruvate oxidase from Escherichia coli: Isolation and properties of the flavoprotein, Arch. Biochem. Biophys. 116:168–176.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1976

Authors and Affiliations

  • Robert B. Gennis
    • 1
  • Lowell P. Hager
    • 2
  1. 1.Departments of Chemistry and BiochemistryUniversity of IllinoisUrbanaUSA
  2. 2.Department of BiochemistryUniversity of IllinoisUrbanaUSA

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