Abstract
Pyruvate kinase (ATP: pyruvate phosphotransferase (EC 2.7.1.40) was partially purified from both autotrophically and heterotrophycally grown Paracoccus denitrificans. The organism grown under heterotrophic conditions contains four times more pyruvate kinase than under autotrophic conditions. The enzyme isolated from both sources exhibited sigmoidal kinetics for both phosphoenolpyruvate (PEP) and ADP. The apparent M m for ADP and PEP in the “autotrophic” enzyme were 0.63 mM ADP and 0.25 mM PEP. The effect of several low molecular weight metabolites on the pyruvate kinase activity was investigated. Ribose-5-phosphate, glucose-6-phosphate and AMP stimulated the reaction at low ADP levels; this stimulation was brought about by an alteration in the apparent K m for ADP. The pyruvate kinases differ in their response to adenine nucleotides, but both preparations seem to be under adenylate control. The results are discussed in relation to the role of pyruvate kinase as a regulatory enzyme in P. denitrificans grown under both autotrophic and heterotrophic conditions.
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Abbreviations
- PEP:
-
phosphoenolpyruvate
- R-5-P:
-
ribose-5-phosphate
- G-6-P:
-
glucose-6-phosphate
- F-6-P:
-
fructose-6-phosphate
- 3-PGA:
-
3-phosphoglycerate
References
Bailey, J.: Techniques in protein chemistry (J. L. Bailey, ed.), pp. 293. Amsterdam: Elsevier 1962
Diesterhaft, M., Freese, E.: Pyruvate kinase of Bacillus subtilis Biochim. Biophys. Acta 268, 373–380 (1972)
Dudouet, D., Le Cam-saginez, M., Bernard, S.: Purification et propriétés cinéfiques de la pyruvate-kinase de Mycobacterium phlei. Biochimie 55, 637–642 (1973)
Forget, P., Pichinoty, F.: Le cycle tricarboxylique chez une bactérie denitrifiante obligatoire. Ann. Inst. Pasteur, Paris 108, 364–377 (1965)
Knobloch, K., Ishaque, M., Aleem, M. I. H.: Oxidative phosphorylation in Micrococcus denitrificans under autotrophic growth conditions. Arch. Mikrobiol. 76, 114–125 (1971)
Kornberg, H. L.: The role and control of the glyoxylate cycle in Escherichia coli. Biochem. J. 99, 1–11 (1966)
Seubert, W., Schoner, W.: The regulation of pyruvate kinase. In: Current topics in cellular regulation, Vol. 3, pp. 237–267 (B. L. Horecker, E. R. Stadtman, eds.) New York-London: Academic Press 1971
Slabas, A. R., Whatley, F. R.: Regulation of pyruvate kinase in Micrococcus denitrificans. Biochem. Soc. Trans. 2, 929–930 (1974)
Slabas, A. R., Whatley, F. R.: Metabolic regulation of the glucose-6-phosphate dehydrogenase from Paracoccus denitrificans grown on glucose/nitrate. Arch. Microbiol. 112, 225–227 (1977)
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Slabas, A.R., Whatley, F.R. Metabolic regulation of pyruvate kinase isolated from autotrophically and heterotrophically grown Paracoccus denitrificans . Arch. Microbiol. 115, 67–71 (1977). https://doi.org/10.1007/BF00427847
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DOI: https://doi.org/10.1007/BF00427847