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Archives of Microbiology

, Volume 159, Issue 2, pp 174–181 | Cite as

Purification of glutaryl-CoA dehydrogenase from Pseudomonas sp., an enzyme involved in the anaerobic degradation of benzoate

  • Ulrich Härtel
  • Elke Eckel
  • Jürgen Koch
  • Georg Fuchs
  • Dietmar Linder
  • Wolfgang Buckel
Original Papers

Abstract

Cell-free extracts of Pseudomonas sp. strains KB 740 and K 172 both contained high levels of glutaryl-CoA dehydrogenase when grown anaerobically on benzoate or other aromatic compounds and with nitrate as electron acceptor. These aromatic compounds have in common benzoyl-CoA as the central aromatic intermediate of anerobic metabolism. The enzymatic activity was almost absent in cells grown aerobically on benzoate regardless whether nitrate was present. Glutaryl-CoA dehydrogenase activity was also detected in cell-free extracts of Rhodopseudomonas, Rhodomicrobium and Rhodocyclus after phototrophic growth on benzoate. Parallel to the induction of glutaryl-CoA dehydrogenase as measured with ferricenium ion as electron acceptor, an about equally high glutaconyl-CoA decarboxylase activity was detected in cell-free extracts. The latter activity was measured with the NAD-dependent assay, as described for the biotin-containing sodium ion pump glutaconyl-CoA decarboxylase from glutamate fermenting bacteria. Glutaryl-CoA dehydrogenase was purified to homogeneity from both Pseudomonas strains. The enzymes catalyse the decarboxylation of glutaconyl-CoA at about the same rate as the oxidative decarboxylation of glutaryl-CoA. The green enzymes are homotetramers (m=170 kDa) and contain 1 mol FAD per subunit. No inhibition was observed with avidin indicating the absence of biotin. The N-terminal sequences of the enzymes from both strains are similar (65%).

Key words

Glutaryl-CoA dehydrogenase Glutaconyl-CoA decarboxylase Pseudomonas sp. Phototrophic proteobacteria Anaerobic degradation of benzoate FAD Ferricenium 

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Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Ulrich Härtel
    • 1
  • Elke Eckel
    • 1
  • Jürgen Koch
    • 2
  • Georg Fuchs
    • 2
  • Dietmar Linder
    • 3
  • Wolfgang Buckel
    • 1
  1. 1.Laboratorium für Mikrobiologie, Fachbereich BiologiePhilipps-UniversitätMarburgFederal Republic of Germany
  2. 2.Angewandte MikrobiologieUniversität UlmUlmFederal Republic of Germany
  3. 3.Biochemisches Institut, Fachbereich HumanmedizinJustus-Liebig-UniversitätGiessenFederal Republic of Germany

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