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. 1964 Jan;87(1):104–113. doi: 10.1128/jb.87.1.104-113.1964

FORMATE—PYRUVATE EXCHANGE REACTION IN STREPTOCOCCUS FAECALIS II.

Reaction Conditions for Cell Extracts

M O Oster a,1, N P Wood a,2
PMCID: PMC276968  PMID: 14102842

Abstract

Oster, M. O. (A. & M. College of Texas, College Station), and N. P. Wood. Formate-pyruvate exchange reaction in Streptococcus faecalis. II. Reaction conditions for cell extracts. J. Bacteriol. 87:104–113. 1964.—In contrast to intact cells of Streptococcus faecalis, no stimulation of the formate-pyruvate exchange reaction was observed in cell extracts when yeast extract was added to the reaction mixture. A heated extract of Micrococcus lactilyticus, vitamin K5, ferrous sulfate, and ferrous ammonium sulfate stimulated an active exchange by protecting the system from oxygen. Tetrahydrofolate, 2,3-dimercaptopropanol, and sodium sulfide provided partial protection, whereas ascorbate, glutathione, sodium hydrosulfite, ammonium sulfide, and sodium bisulfite gave insufficient protection or were inhibitory. Oxidation-reduction (O-R) indicators were not inhibitory and were used to estimate the O-R potentials of reaction mixtures. A potential at least as negative as −125 mv was estimated to be necessary to preserve or initiate formate-pyruvate exchange activity. The reaction operated over a narrow pH range when strict anaerobic conditions were not maintained but, when the system was suitably poised, the pH range was broader. The influence of high phosphate concentrations was less under strictly anaerobic conditions, and orthophosphate could be replaced by small amounts of pyrophosphate. Effect of temperature, time, and amount of extract is presented. Addition of reduced benzyl viologen and hydrogen-saturated palladium in the buffer during 8 hr of dialysis prevented inactivation of extracts. Recovery of activity could be obtained after ammonium sulfate treatment when a combination of palladium chloride, neutral red, and hydrogen bubbling were used.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Gunsalus I. C., Campbell J. J. Diversion of the Lactic Acid Fermentation with Oxidized Substrate. J Bacteriol. 1944 Oct;48(4):455–461. doi: 10.1128/jb.48.4.455-461.1944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  3. MORTLOCK R. P., VALENTINE R. C., WOLFE R. S. Carbon dioxide activation in the pyruvate clastic system of Clostridium butyricum. J Biol Chem. 1959 Jul;234(7):1653–1656. [PubMed] [Google Scholar]
  4. McCormick N. G., Ordal E. J., Whiteley H. R. DEGRADATION OF PYRUVATE BY MICROCOCCUS LACTILYTICUS II. : Studies of Cofactors in the Formate-Exchange Reaction. J Bacteriol. 1962 Apr;83(4):899–906. doi: 10.1128/jb.83.4.899-906.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. NOVELLI G. D. The exchange of H14COOH with the carboxyl group of pyruvate by Clostridium butylicum and Micrococcus lactilyticus. Biochim Biophys Acta. 1955 Dec;18(4):594–596. doi: 10.1016/0006-3002(55)90170-0. [DOI] [PubMed] [Google Scholar]
  6. O'kane D. J., Gunsalus I. C. Pyruvic Acid Metabolism: A Factor Required for Oxidation by Streptococcus faecalis. J Bacteriol. 1948 Oct;56(4):499–506. doi: 10.1128/jb.56.4.499-506.1948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. PECK H. D., Jr, GEST H. Formic dehydrogenase and the hydrogenlyase enzyme complex in coli-aerogenes bacteria. J Bacteriol. 1957 Jun;73(6):706–721. doi: 10.1128/jb.73.6.706-721.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. RABINOWITZ J. C., ALLEN E. H. Role of vitamin B12 derivatives in reactions of pyruvate. Fed Proc. 1961 Dec;20:962–966. [PubMed] [Google Scholar]
  9. STRECKER H. J. Formate fixation in pyruvate by Escherichia coli. J Biol Chem. 1951 Apr;189(2):815–830. [PubMed] [Google Scholar]
  10. WHITELEY H. R., DOUGLAS H. C. The fermentation of purines by Micrococcus lactilyticus. J Bacteriol. 1951 May;61(5):605–616. doi: 10.1128/jb.61.5.605-616.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. WHITELEY H. R., McCORMICK N. G. Degradation of pyruvate by Micrococcus lactilyticus. III. Properties and cofactor requirements of the carbon dioxide-exchange reaction. J Bacteriol. 1963 Feb;85:382–393. doi: 10.1128/jb.85.2.382-393.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. WOLFE R. S., O'KANE D. J. Cofactors of the phosphoroclastic reaction of Clostridium butyricum. J Biol Chem. 1953 Dec;205(2):755–765. [PubMed] [Google Scholar]
  13. WOOD N. P., O'KANE D. J. FORMATE-PYRUVATE EXCHANGE REACTION IN STREPTOCOCCUS FAECALIS. I. FACTOR REQUIREMENT FOR INTACT CELLS. J Bacteriol. 1964 Jan;87:97–103. doi: 10.1128/jb.87.1.97-103.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]

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