Summary
The assimilation of14CO2 and [2-14C] acetate, [3-14C] pyruvate, [5-14C] α-ketoglutarate, [2,3-14C] succinate, [U-14C] glutamate and [U-14C] aspartate was followed in cell suspensions ofNitrosomonas europaea andNitrobacter agilis respectively. There was appreciable incorporation of these substrates even without adding the inorganic nitrogen compounds that are oxidized by these bacteria yielding ATP. In the soluble amino acid fraction most of14C label was recovered in glutamate while in the protein amino acids a more uniform distribution was found. Acetate was rapidly incorporated to a high level in both nitrifying bacteria while inNitrobacter there was a relatively lower uptake of the other substrates especially succinate. High levels of the NAD malate dehydrogenase and NADP isocitrate dehydrogenase were measured but no significant amounts of the other tricarboxylic acid cycle enzymes or NADH oxidase were found. Glutamate decarboxylase was detected in both organisms and the transferase assay for glutamine synthetase indicated a 30-fold higher activity for this enzyme inNitrobacter. The amino acid composition of the water soluble fraction was determined in both bacteria.
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References
Aleem, M. I. H.: Path of carbon and assimilatory power in chemosynthetic bacteria. I.Nitrobacter agilis. Biochim. blophys. Acta (Amst.)107, 14–28 (1965).
—: Mechanism of oxidative phosphorylation in the chemoautotrophNitrobacter agilis. Biochim. biophys. Acta (Amst.)162, 338–347 (1968).
—, Alexander, M.: Cell-free nitrification byNitrobacter. J. Bact.76, 510–514 (1958).
—, Lees, H., Nicholas, D. J. D.: Adenosine triphosphate-dependent reduction of nicotinamide adenine dinucleotide by ferro-cytochrome c in chemoautotrophic bacteria. Nature (Lond.)200, 759–761 (1963).
Amarasingham, C. R., Davis, B. D.: Regulation of α-ketoglutarate dehydrogenase formation inEscherichia coli. J. biol. Chem.240, 3664–3668 (1965).
Anderson, K. J., Lundgren, D. C.: Enzymatic studies of the iron oxidizing bacteriumFerrobacillus ferrooxidans: evidence for a glycolytic pathway and Krebs cycle. Canad. J. Microbiol.15, 73–79 (1969).
Bernath, P., Singer, T. P.: Succinic dehydrogenase. In: Methods in Enzymology, Vol. 5, pp. 597–614. Ed. by S. P. Colowick and N. O. Kaplan. New York: Academic Press, Inc. 1962.
Boulter, D., Barber, J. T.: Amino acid metabolism in germinating seeds ofVicia faba L. in relation to their biology. New Phytol.62, 301–316 (1963).
Butler, R. G., Umbreit, W. W.: Reduced nicotinamide adenine dinucleotide oxidase and α-ketoglutaric dehydrogenase activity byThiobacillus thiooxidans. J. Bact.97, 966–967 (1969).
Campbell, A. E., Hellebust, J. A., Watson, S. W.: Reductive pentose phosphate cycle inNitrosocystis oceanus. J. Bact.91, 1178–1185 (1966).
Clark, C., Schmidt, E. L.: Effect of mixed culture onNitrosomonas europaea simulated by uptake and utilization of pyruvate. J. Bact.91, 367–373 (1966).
——: Growth response ofNitrosomonas europaea to amino acids. J. Bact.93, 1302 to 1308 (1967a).
——: Uptake and utilization of amino acids by resting cells ofNitrosomonas europaea. J. Bact.93, 1309–1315 (1967b).
Delwiche, C. C., Finstein, M. S.: Carbon and energy sources for the nitrifying autotrophNitrobacter. J. Bact.90, 102–107 (1965).
Dixon, G. H., Kornberg, H. L.: Assay methods for key enzymes of the glyoxylate cycle. Biochem. J.72, 3 P (1959).
Evans, M. C. W., Buchanan, B. B., Arnon, D. I.: A new ferredoxin-dependent carbon reduction cycle in a photosynthetic bacterium. Proc. nat. Acad. Sci. (Wash.)55, 928–934 (1966).
Faull, K. F., Wallace, W., Nicholas, D. J. D.: Nitrite oxidase and nitrate reductase inNitrobacter agilis. Biochem. J.113, 449–455 (1969).
Fonnum, F.: The distribution of glutamate decarboxylase and aspartate transaminase in sub-cellular fractions of rat and guinea pig brain. Biochem. J.106, 401–412 (1968).
Grivell, A. R., Jackson, J. F.: Thymidine kinase: Evidence for its absence fromNeurospora crassa and some other micro-organisms, and the relevance of this to the specific labelling of deoxyribonucleic acid. J. gen. Microbiol.54, 307–317 (1968).
Hager, L. P., Kornberg, H. L.: On the mechanism of α-oxoglutarate oxidation inEscherichia coli. Biochem. J.78, 194–198 (1961).
Harris, G. K., Tigane, E., Hanes, G. S.: Quantitative chromatographic methods. 7. Isolation of amino acids from serum and other fluids. Canad. J. Biochem. Physiol.39, 439–451 (1961).
Hoare, D. S., Hoare, S. L., Moore, R. B.: The photoassimilation of organic compounds by autotrophic blue-green algae. J. gen. Microbiol.49, 351–370 (1967).
Hofman, T.: The biochemistry of the nitrifying organisms. 3. Composition ofNitrosomonas. Biochem. J.54, 293–295 (1953).
Hooper, A. B.: Biochemical basis of obligate autotrophy inNitrosomonas europaea. J. Bact.97, 776–779 (1969).
Ida, S., Alexander, M.: Permeability ofNitrobacter agilis to organic compounds. J. Bact.90, 151–156 (1965).
Jennings, A. C., Morton, R. K.: Amino acids and protein synthesis in developing wheat endosperm. Aust. J. biol. Sci.16, 384–394 (1963).
John, P. C. L., Syrett, P. J.: The purification and properties of isocitrate lyase fromChlorella. Biochem. J.105, 409–416 (1967).
Johnson, E. J., Peck, H. D. Jr: Coupling of phosphorylation and carbon dioxide fixation in extracts ofThiobacillus thioparus. J. Bact.89, 1041–1050 (1965).
Kaufman, S.: α-Ketoglutaric dehydrogenase system and phosphorylating enzyme from heart muscle. In: Methods in Enzymology, Vol.1, pp. 714–722. Ed. by S. P. Colowick and N. O. Kaplan. New York: Academic Press, Inc. 1955.
Kelly, D. P.: The incorporation of acetate by the chemoautotrophThiobacillus neapolitanus, Strain C. Arch. Mikrobiol.58, 99–115 (1967).
Leach, C. K., Carr, N. G.: Reduced nicotinamide-adenine dinucleotide phosphate oxidase in the autotrophic blue-green algaAnabaena variabilis. Biochem. J.109, 4P (1968).
Lees, H.: Symposium on autotrophy. IV. Some thoughts on the energetics of chemosynthesis. Bact. Rev.26, 165–167 (1962).
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. biol. Chem.193, 265–275 (1951).
Machlis, L., Torrey, J. G.: Plants in action, p. 44. San Francisco: W. H. Freeman and Company 1956.
Mahler, H. R., Cordes, E. H., Biological chemistry. pp. 441–442. New York: Harper and Row Publ. Inc. 1966.
Meister, A.: Biochemistry of amino acids. New York: Academic Press, Inc. 1965.
Moon, K. E., Schofield, P. J.: Utilization of tricarboxylic acid cycle intermediates byHaemonchus contortus larvae. Comp. Biochem. Physiol.24, 581–590 (1968).
Murray, R. G. E., Watson, S. W.: Structure ofNitrosocystis oceanus and comparison withNitrosomonas andNitrobacter. J. Bact.89, 1594–1609 (1965).
Nicholas, D. J. D., Rao, P. S.: The incorporation of labelled CO2 into cells and extracts ofNitrosomonas europaea. Biochim. biophys. Acta (Amst.)82, 394–397 (1964).
Oaks, A.: The soluble leucine pool in maize root tips. Plant Physiol.40, 142–149 (1965).
Ochoa, S.: Isocitric dehydrogenase system (NADP+) from pig heart. In: Methods in Enzymology, Vol. 1, pp. 699–707. Ed. by S. P. Colowick and N. O. Kaplan. New York: Academic Press, Inc. 1955a).
—: Malic dehydrogenase from pig heart. In: Methods in Enzymology, Vol. 1, pp. 735–739. Ed. by S. P. Colowick and N. O. Kaplan. New York: Academic Press, Inc. 1955b).
Pate, J. S., Walker, J., Wallace, W.: Nitrogen containing compounds in the shoot system inPisum arvense L. II. The significance of the amino-acids and amides released from nodulated roots. Ann. Bot., (Lond.) N. S.29, 475–493 (1965).
Polakis, E. S., Bartley, W.: Changes in the enzyme activities ofSaccharomyces cerevisiae during aerobic growth on different carbon sources. Biochem. J.97, 284–297 (1965).
Racker, E.: Spectrophotometric measurements of the enzymatic formation of fumaric and cis-aconitic acids. Biochim. biophys. Acta (Amst.)4, 211–214 (1950).
Rao, P. S., Nicholas, D. J. D.: Studies on the incorporation of CO2 by cells and cell-free extracts ofNitrosomonas europaea. Biochim. biophys. Acta (Amst.)124, 221–232 (1966).
Roberts, R. B., Cowie, D. B., Abelson, P. H., Bolton, E. J., Britten, R. J.: Studies of biosynthesis inEscherichia coli. Carnegie Institution of Washington, Publication, No. 607 (1955).
Simmonds, D. H., Rowlands, R. J.: Automatic equipment for simultaneous determination of amino acids separated on several ion exchange resin columns. Analyt. Chem.32, 259–268 (1960).
Smith, I.: Chromatographic and electrophoretic techniques. pp. 6–39. London: Heinemann 1960.
Smith, A. J., Hoare, D. S.: Acetate assimilation byNitrobacter agilis in relation to its obligate autotrophy. J. Bact.95, 844–855 (1968).
—, London, J., Stainer, R. Y.: Biochemical basis of obligate autotrophy in bluegreen algae andThiobacilli. J. Bact.94, 972–983 (1967).
Tobback, P., Laudelout, H.: Poly-β-hydroxybutyric acid inNitrobacter. Biochim. biophys. Acta (Amst.)97, 589–590 (1965).
Trudinger, P. A., Kelly, D. P.: Reduced nicotinamide adenine dinucleotide oxidation byThiobacillus neapolitanus andThiobacillus strain c. J. Bact.95, 1962 to 1963 (1968).
Wallace, W., Nicholas, D. J. D.: Properties of some reductase enzymes in the nitrifying bacteria and their relationship to the oxidase systems. Biochem. J.109, 763–773 (1968).
——: Glutamate dehydrogenase inNitrosomonas europaea and the effect of hydroxylamine, oximes and related compounds on its activity. Biochim. biophys. Acta (Amst.)171, 229–237 (1969).
Williams, J. LeB., Watson, S. W.: Autotrophy inNitrosocystis oceanus. J. Bact.96, 1640–1648 (1968).
Woolfolk, C. A., Shapiro, B., Stadtman, E. R.: Regulation of glutamine synthetase. I. Purification and properties of glutamine synthetase fromEscherchia coli. Arch. Biochem116, 177–192 (1966).
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Wallace, W., Knowles, S.E. & Nicholas, D.J.D. Intermediary metabolism of carbon compounds by nitrifying bacteria. Archiv. Mikrobiol. 70, 26–42 (1970). https://doi.org/10.1007/BF00691058
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DOI: https://doi.org/10.1007/BF00691058