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Archives of Microbiology

, Volume 110, Issue 2–3, pp 157–166 | Cite as

Purification, some properties and quaternary structure of thed-ribulose 1,5-diphosphate carboxylase ofAlcaligenes eutrophus

  • B. Bowien
  • F. Mayer
  • G. A. Codd
  • H. G. Schlegel
Article

Abstract

d-Ribulose 1,5-diphosphate carboxylase has been purified from autotrophically grown cells of the facultative chemolithotrophic hydrogen bacteriumAlcaligenes eutrophus. The enzyme was homogeneous by the criteria of polyacrylamide gel electrophoresis. The molecular weight of the enzyme was 505000 determined by gel filtration and sucrose density gradient centrifugation, and a sedimentation coefficient of 18.2 S was obtained. It was demonstrated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis that the enzyme consists of two types of subunits of molecular weight 52000 and 13000.

Electron microscopy on the intact and the partially dissociated enzyme lead to the construction of a model for the quaternary structure of the enzyme which is composed of 8 large and 8 small subunits. The most probable symmetry of the enzyme molecule is 4:2:2.

Michaelis constant (K m ) values for ribulose 1,5-diphosphate, Mg2-, and CO2 were 0.59 mM, 0.33 mM, and 0.066 mM measured under air. Oxygen was a competitive inhibitor with respect to CO2 suggesting that the enzyme also exhibits an oxygenase activity. The oxygenolytic cleavage of ribulose 1,5-diphosphate was shown and a 1:1 stoichiometry between oxygen consumption and 3-phosphoglycerate formation observed.

Key words

d-Ribulose 1,5-diphosphate carboxylase Oxygenase activity Quaternary structure Electron microscopy Alcaligenes eutrophus 

Abbreviations

DTE

dithioerythritol

EDTA

ethylenediamine tetraacetate

RuDP

d-ribulose 1,5-diphosphate

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Copyright information

© Springer-Verlag 1976

Authors and Affiliations

  • B. Bowien
    • 1
    • 2
  • F. Mayer
    • 1
    • 2
  • G. A. Codd
    • 1
    • 2
  • H. G. Schlegel
    • 1
    • 2
  1. 1.Institut für Mikrobiologie der UniversitätGöttingenFederal Republic of Germany
  2. 2.der Gesellschaft für Strahlen-und Umweltforschung mbHGöttingenFederal Republic of Germany

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