Abstract
Acetate uptake by strains of Synechococcus and Aphanocapsa in short experiments required light, and was strongly inhibited by m-dichlorocarbonyl cyanide phenylhydrazone and dichlorophenyl dimethyl urea. Acetate carbon was distributed in amino acids and in the acyl portion of lipids in the same way as during growth experiments when CO2 was available, but the reduced incorporation in the absence of CO2 was primarily into the lipid fraction. An apparent K m for uptake by Synechococcus and for Aphanocapsa 6308 of 20 and 180 μM at pH 7.4 was obtained; corresponding V max values were 6 and 11 nmol x min-1 x mg protein-1. Uptake with Synechococcus was affected by pH, with affinity decreased and maximal rate increase with rising pH. Acetate uptake was not affected by propionate or butyrate when both were added at the same time, but a light and concentration dependent inhibition developed if suspensions were preincubated with propionate. Acetate carbon moved rapidly into acid insoluble material, but after 10–15 s 75% or more of the recovered intracellular counts were in acetyl CoA. Counts in this compound were reduced by preincubation with propionate.
Kinetic measurements of acetyl CoA synthetase in fractionated cell extracts gave values for K m of about 50 μM for acetate, 5 mM for propionate, 100 μM for CoA and 0.38 mM for ATP. The internal pool of free CoA was measured to be about 20 μM, and was reduced by preincubation with propionate. This suggests that the activity of CoA-mediated reactions may be regulated by the availability of this cofactor.
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Abbreviations
- CCCP:
-
m-Dichlorocarbonyl cyanide phenyl hydrazone
- DCMU:
-
dichlorophenyl dimethyl urea
- TCA:
-
trichloroacetic acid
- Tris:
-
trishydroxymethyl amino methane
- HEPES:
-
N-2-hydroxyethylpiperazine-N′-2-ethane-sulfonic acid
References
Allred, J. B., Guy, D. G.: Determination of coenzyme A and acetyl CoA in tissue extracts. Analyt. Biochem. 29, 293–299 (1969)
Biggins, J.: Respiration in blue-green algae. J. Bact. 99, 570–575 (1969)
Bothe, H., Nolteernsting, U.: Pyruvate dehydrogenase complex, pyruvate: ferredoxin oxidoreduction and lipoic acid content of microorganisms. Arch. Microbiol. 102, 53–57 (1975)
Carr, N. G.: Metabolic control and autotrophic physiology. In: The biology of the blue-green algae (N. G. Carr, B. A. Whitton, eds.). Berkeley-Los Angeles: Univ. of Calif. Press 1973
Cocking, E. C., Yemm, E.: Estimation of amino acids by ninhydrin. Biochem. J. 58, xii (1954)
Frerman, F. E.: The role of acetyl CoA: butyrate coenzyme A transferase in the uptake of butyrate by isolated membrane vesicles of E. coli. Arch. Biochem. Biophys. 159, 444–452 (1973)
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)
Hoare, D. S., Moore, R. B.: Photoassimilation of organic compounds by autotrophic blue-green algae. Biochim. biophys. Acta (Amst.) 109, 622–625 (1965)
Holm, H. W., Vennes, J. W.: Occurrence of purple sulfur bacteria in a sewage treatment lagoon. Appl. Microbiol. 19, 988–996 (1970)
Ihlenfeldt, M. J. A., Gibson, J.: CO2 fixation and its regulation in Anacystis nidulans (Synechococcus). Arch. Microbiol. 102, 13–22 (1975a)
Ihlenfeldt, M. J. A., Gibson, J.: Phosphate utilization and alkaline phosphatase in Anacystis nidulans (Synechococcus). Arch. Microbiol. 102, 23–28 (1975b)
Jones, M. E., Lipmann, F.: Aceto-CoA-kinase. In: Methods in enzymology (S. P. Colowick, N. O. Kaplan, eds.), Vol. 1, pp. 585–591. New York: Academic Press 1955
Kelly, D. P.: Autotrophy: concepts of lithotrophic bacteria and their organic metabolism. Ann. Rev. Microbiol. 25, 177–210 (1971)
Law, J. H., Slepecky, R. A.: Assay of poly-β-hydroxybutyric acid. J. Bact. 82, 33–36 (1961)
Lowry, O. H., Rosebrough, A. L., Farr, A. L., Randall, R. J.: Protein measurement with the Folin reagent. J. biol. Chem. 193, 265–275 (1951)
Pelroy, R. A., Bassham, J. A.: Photosynthetic and dark carbon metabolism in unicellular blue-green algae. Arch. Mikrobiol. 86, 25–38 (1972)
Pelroy, R. A., Rippka, R., Stanier, R. Y.: Metabolism of glucose by unicellular blue-gree algae. Arch. Mikrobiol. 87, 303–322 (1972)
Rittenberg, S. C.: The role of exogeneous organic matter in the physiology of chemolithotrophic bacteria. In: Adv. microbial physiol. (A. H. Rose, J. H. Wilkinson, eds.), Vol. 3, pp. 159–196. New York: Academic Press 1969
Roberts, R. B.: Abelson, P. H., Cowie, D. B., Bolton, E. T., Britton, R. J.: Studies of biosynthesis in Escherichia coli. Carnegie Institute, Washington (1957)
Saunders, G. W.: Potential heterotrophy in a natural population of Oscillatoria agardhii var isothrix Skuja. Limnol. Oceanogr. 17, 704–711 (1972)
Scholes, P., Mitchell, P., Moyle, J.: The polarity of proton translocation in some photosynthetic microorganisms. Europ. J. Biochem. 8, 450–454 (1969)
Simon, E. J., Shemin, D.: Preparation of coenzyme A derivatives. J. Amer. chem. Soc. 75, 2520 (1953)
Smith, A. J., London, J., Stanier, R. Y.: Biochemical basis of obligate autotrophy in blue-green algae and thiobacilli. J. Bact. 94, 972–983 (1967)
Smith, A. J., Lucas, C.: Propionate resistance in blue-green algae. Biochem. J. 124, 23P (1971)
Tokumitsu, Y., Ui, M.: A new simple technique for the determination of radioactivity content of coenzyme A products. Analyt. Biochem. 59, 1780–1784 (1974)
Wagner, B. J., Miović, M. L., Gibson, J.: Utilization of amino acids by Chromatium sp., Strain, D. Arch. Mikrobiol. 91, 255–272 (1973)
White, D. C., Frerman, F. E.: Extraction, characterization and cellular localization of lipids of Staphylococcus aureus. J. Bact. 94, 1854–1867 (1967)
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Ihlenfeldt, M.J.A., Gibson, J. Acetate uptake by the unicellular cyanobacteria Synechococcus and Aphanocapsa . Arch. Microbiol. 113, 231–241 (1977). https://doi.org/10.1007/BF00492030
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DOI: https://doi.org/10.1007/BF00492030