Archiv für Mikrobiologie

, Volume 59, Issue 1–3, pp 4–12 | Cite as

Citramalate lyase of Clostridium tetanomorphum

  • H. A. Barker


Assay methods and some properties of (+)-citramalate pyruvatelyase, an enzyme from Clostridium tetanomorphum that converts (+)-citramalate to pyruvate and acetate, are described. The enzyme is very active (0.8–1.2 units per mg protein) in freshly prepared extracts, but loses activity rapidly during storage. (+)-Citramalate is the only substrate found to be cleaved by the lyase; the equilibrium for the reaction permits almost complete cleavage at low substrate concentrations. A divalent cation is required as a cofactor. A sensitive and specific enzymic method for estimating (+)-citramalate is described.


Enzyme Acetate Pyruvate Substrate Concentration Clostridium 
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  1. Barker, H. A.: Separation, identification, and estimation of lower fatty acids. In: Methods in Enzymology (Colowick, S. P., and N. O. Kaplan, eds.), vol. III, pp. 372–382. New York: Academic Press 1957.Google Scholar
  2. —: Fermentation of nitrogenous organic compounds. In: The Bacteria (Gunsalus, I. C., and R. Y. Stanier, eds.), vol. II, pp. 151–207. New York: Academic Press 1961.Google Scholar
  3. —: Chemical synthesis and resolution of (±)-citramalic acid. In: Biochemical Preparations (Coon, M. J., ed.), vol. 9, pp. 25–29. New York: J. Wiley 1962.Google Scholar
  4. —, and A. H. Blair: Enzymatic synthesis of (+)-citramalic acid. In: Biochemical Preparations (Coon, M. J., ed.), vol. 9, pp. 21–25. New York: J. Wiley 1962.Google Scholar
  5. —, V. Rooze, F. Suzuki, and A. A. Iodice: The glutamate mutase system. Assays and properties. J. biol. Chem. 239, 3260–3266 (1964).Google Scholar
  6. —, R. D. Smyth, R. M. Wilson, and H. Weissbach: The purification and properties of β-methylaspartase. J. biol. Chem. 234, 320–328 (1959).Google Scholar
  7. Barker, S. B., and W. H. Summerson: The colorimetric determination of lactic acid in biological material. J. biol. Chem. 138, 535–554 (1941).Google Scholar
  8. Benedict, C. R.: Early products of [14C]acetate incorporation in resting cells of Rhodospirillum rubrum. Biochim. biophys. Acta (Amst.) 56, 620–622 (1962).Google Scholar
  9. Blair, A. H., and H. A. Barker: Assay and purification of (+)-citramalate hydrolyase components of Clostridium tetanomorphum. J. biol. Chem. 241, 400–408 (1966).Google Scholar
  10. Friedemann, T. E., and G. E. Haugen: Pyruvic acid. II. The determination of keto acids in blood and urine. J. biol. Chem. 147, 415–442 (1943).Google Scholar
  11. Gray, C. T., and H. L. Kornberg: Enzymatic formation of citramalate from acetylcoenzyme A and pyruvate in Pseudomonas ovalis Chester, catalysed by “pyruvate transacetase”. Biochim. biophys. Acta (Amst.) 42, 371–372 (1960).Google Scholar
  12. Hulme, A. C.: The isolation of L-citramalic acid from the peel of the apple fruit. Biochim. biophys. Acta (Amst.) 14, 36–43 (1954).Google Scholar
  13. Kennedy, E. P., and H. A. Barker: Paper chromatography of volatile acids. Analyt. Chem. 23, 1033–1034 (1951).Google Scholar
  14. Kinnory, D. S., Y. Takeda, and D. M. Greenberg: Chromatography of carboxylic acids on a silica gel column with a benzene-ether solvent system. J. biol. Chem. 212, 379–396 (1955).Google Scholar
  15. Kornberg, A.: Lactic dehydrogenase of muscle. In: Methods in Enzymology (Colowick, S. P., and N. O. Kaplan, eds.), vol. I, pp. 441–443. New York: Academic Press 1955.Google Scholar
  16. Losada, M., A. V. Trebst, S. Ogata, and D. I. Arnon: Equivalence of light and adenosine triphosphate in bacterial photosynthesis. Nature (Lond.) 186, 753–760 (1960).Google Scholar
  17. Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall: Protein measurement with the folin phenol reagent. J. biol. Chem. 193, 265–275 (1951).Google Scholar
  18. Nagai, J.: Studies on itaconate metabolism. II. Citramalate metabolism in Pseudomonas fluorescens growth on itaconate, J. Biochem. (Tokyo) 54, 34–40 (1963).Google Scholar
  19. Smith, R. A., J. R. Stamer, and I. C. Gunsalus: Citritase and isocitritase reactions: Equilibria-energetics. Biochim. biophys. Acta (Amst.) 19, 567–568 (1956).Google Scholar

Copyright information

© Springer-Verlag 1967

Authors and Affiliations

  • H. A. Barker
    • 1
  1. 1.Department of BiochemistryUniversity of CaliforniaBerkeleyUSA

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