Abstract
The formate dehydrogenases of Clostridium acidiurici and of C. cylindrosporum coupled the oxidation of formate with the reduction of viologen dyes. The basal activity level was about 0.85 μmoles/min s mg of protein for both species. The level of formate dehydrogenase of C. acidiurici increased 12-fold when 10-7 M tungstate and selenite were present during growth. Molybdate exerted no effect. On the other hand, molybdate and selenite were required to increase the formate dehydrogenase of C. cylindrosporum, and tungstate exhibited an antagonistic effect in this organism.
Growth on hypoxanthine generally depended on the addition of bicarbonate. Supplementation with tungstate and selenite accelerated growth of C. acidiurici and increased again the level of formate dehydrogenase. The addition of both, molybdate and selenite was necessary to initiate growth of C. cylindrosporum and to form an active formate dehydrogenase.
The differences in the requirement for metal ion supplementation to form high levels of formate dehydrogenase and their involvement in hypoxanthine degradation can be used to differentiate between C. acidiurici and C. cylindrosporum.
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Abbreviations
- FDH:
-
formate dehydrogenase
References
Andreesen, J. R., Gottschalk, G., Schlegel, H. G.: Clostridium formicoaceticum nov. spec. Isolation, description and distinction from C. aceticum and C. thermoaceticum. Arch. Mikrobiol 72, 154–174 (1970)
Andreesen, J. R., Ljungdahl, L. G.: Formate dehydrogenase of Clostridium thermoaceticum: incorporation of selenium-75, and the effect of selenite, molybdate, and tungstate on the enzyme. J. Bacteriol. 116, 867–873 (1973)
Andreesen, J. R., Ljungdahl, L. G.: Nicotinamide adenine dinucleotide phosphate-dependent formate dehydrogenase from Clostridium thermoaceticum: Purification and properties. J. Bacteriol. 120, 6–14 (1974)
Andreesen, J. R., El Ghazzawi, E., Gottschalk, G.: The effects of ferrous ions, tungstate and selenite on the level of formate dehydrogenase in Clostridium formicoaceticum and formate synthesis from CO2 during pyruvate fermentation. Arch. Microbiol. 96, 103–118 (1974)
Barker, H. A.: Fermentations of nitrogenous organic compounds. In: The bacteria, Vol. II: Metabolism (I. C. Gunsalus, R. Y. Stainer, eds.), pp. 151–207. New York-London: Academic Press 1961
Barker, H. A., Beck, J. V.: The fermentative decomposition of purines by Clostridium acidici-urici and Clostridium cylindrosporum. J. Biol. Chem. 141, 3–27 (1941)
Barker, H. A., Beck, J. V.: Clostridium acidi-urici and Clostridium cylindrosporum, organisms fermenting uric acid and some other purines. J. Bacteriol. 43, 291–304 (1942)
Barker, H. A., Elsden, S. R.: Carbon dioxide utilization in the formation of glycine and acetic acid. J. Biol. Chem. 167, 619–620 (1947)
Beisenherz, G., Bolze, H. J., Bücher, Th., Czok, R., Garbade, K. H., Meyer-Arendt, E., Pfleiderer, G.: Diphosphofructose-Aldolase, Phosphoglyceraldehyd-Dehydrogenase, Milchsäure-Dehydrogenase, Glycerophosphat-Dehydrogenase und Pyruvat-Kinase aus Kaninchenmuskulatur in einem Arbeitsgang. Z. naturforsch. 8b, 555–577 (1953)
Bradshaw, W. H., Barker, H. A.: Purification and properties of xanthine dehydrogenase from Clostridium cylindrosporum. J. Biol. Chem. 235, 3620–3629 (1960)
Bradshaw, W. H., Reeder, D. J.: Ferredoxin coupling of formate oxidation to urate reduction in extracts of Clostridium cylindrosporum. Bacteriol. Proc., p. 110 (1964)
Bray, R. C.: Molybdenum iron-sulfur flavin hydroxylases and related enzymes: In: The enzymes, 3rd ed., Vol. 12 (P. Boyer, ed.), pp. 300–418 New York-London: Academic Press 1975
Corwin, A. H., Arellano, R. R., Chivvis, A. B.: Anomalies of viologens in bases and water. Biochim. Biophys. Acta 162, 533–538 (1968)
Enoch, H. G., Lester, R. L.: Effects of molybdate, tungstate, and selenium compounds on formate dehydrogenase and other enzyme systems in Escherichia coli. J. Bacteriol. 110, 1032–1040 (1972)
Enoch, H. G., Lester, R. L.: The purification and properties of formate dehydrogenase and nitrate reductase from Escherichia coli. J. Biol. Chem. 250, 6693–6705 (1975)
Gottschalk, G., Barker, H. A.: Presence and stereospecificity of citrate synthase in anaerobic bacteria. Biochemistry 6, 1027–1034 (1967)
Karlsson, J. L., Barker, H. A.: Tracer experiments on the mechanism of uric acid decomposition and acetic acid synthesis by Clostridium acidi-urici. J. Biol. Chem. 178, 891–902 (1949)
Kearny, J. J., Sagers, R. D.: Formate dehydrogenase from Clostridium acidiurici. J. Bacteriol. 109, 152–161 (1972)
Ljungdahl, L. G., Andreesen, J. R.: Tungsten, a component of active formate dehydrogenase of Clostridium thermoaceticum. FEBS-Lett. 54, 279–282 (1975)
Ljungdahl, L. G., Andreesen, J. R.: Reduction of CO2 to acetate in homoacetate fermenting clostridia and the involvement of tungsten in formate dehydrogenese. In: Sympsium on microbial production and utilization of gases (H2, CH4, CO) (H. G. Schlegel, G. Gottschalk, N. Pfennig, eds.), pp. 163–172, Göttingen Goltze 1976
Ljungdahl, L. G., Sherod, D. W., Moore, M. R., Andreesen, J. R.: Properties of enzymes from Clostridium thermoaceticum and Clostridium formicoaceticum. Experientia Suppl. 26, 237–248 (1976)
Lovenberg, W., Buchanan, B. B., Raninowitz, J. C.: Studies on the chemical nature of clostridial ferredoxin. J. Biol. Chem. 238, 3899–3913 (1963)
Michaelis, L., Hill, E. S.: The viologen indicators. J. Gen. Physiol. 16, 859–873 (1933)
O'Brien, W. E., Brewer, J. M., Ljungdahl, L. G.: Chemical, physical and enzymatic comparisons of formyltetrahydrofolate synthetases from thermo- and mesophilic clostridia. Experientia Suppl. 26, 249–262 (1976)
Rabinowitz, J. C.: Intermediates in purine breakdown. In: Methods in enzymology, Vol. 6 (S. P. Colowick, N. O. Kaplan, eds.), pp. 703–713 New York-London: Academic Press 1963
Rabinowitz, J. C., Barker, H. A.: Purine fermentation by Clostridium cylindrosporum. II. Purine transformations. J. Biol. Chem. 218, 161–173 (1956)
Rabinowitz, J. C., Himes, R. H.: Folic acid coenzymes. Fed. Proc. 19, 963–970 (1960)
Sagers, R. D., Carter, J. E.: l-Serine dehydratase (Clostridium acidiurici). In: Methods in enzymology, Vol. 17 (B. H. Tabor, C. W. Tabor, eds.), pp. 351–356. New York-London, Academic Press 1971
Schulman, M., Parker, D., Ljungdahl, L. G., Wood, H. G.: Total synthesis of acetate from CO2. V. Determination by mass analysis of the different types of acetate formed from 13CO2 by heterotrophic bacteria. J. Bacteriol. 109, 633–644 (1972)
Stadtman, T. C.: Selenium biochemistry. Science 183, 915–922 (1974)
Thauer, R. K.: CO2-Reduction to formate by NADPH. The initial step in the total synthesis of acetate from CO2 in Clostridium thermoaceticum. FEBS-Lett. 27, 111–115 (1972)
Thauer, R. K.: CO2 reduction to formate in Clostridium acidiurici. J. Bacteriol. 114, 443–444 (1973)
Thauer, R. K., Fuchs, G., Schnitker, U., Jungermann, K.: CO2 reductase from Clostridium pasteurianum: molybdenum dependence of synthesis and inactivation by cyanide. FEBS-Lett. 38, 45–48 (1973)
Thorneley, R. V. F.: A convenient electrochemical preparation of reduced methyl viologen and a kinetic study of the reaction with oxygen using an anaerobic stopped-flow apparatus. Biochim. Biophys. Acta 333, 487–496 (1974)
Vogels, G. D., van der Drift, C.: Degradation of purines and pyrimidines by microorganisms. Bacteriol. Rev. 40, 403–468 (1976)
Wood, H. G.: A study of carbon dioxide fixation by mass determination of the types of C13-acetate. J. Biol. Chem. 194, 905–931 (1952)
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Wagner, R., Andreesen, J.R. Differentiation between Clostridium acidiurici and Clostridium cylindrosporum on the basis of specific metal requirements for formate dehydrogenase formation. Arch. Microbiol. 114, 219–224 (1977). https://doi.org/10.1007/BF00446865
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DOI: https://doi.org/10.1007/BF00446865