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The periplasmic nitrate reductase of Rhodobacter capsulatus; purification, characterisation and distinction from a single reductase for trimethylamine-N-oxide, dimethylsulphoxide and chlorate

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Abstract

The periplasmic dissimilatory nitrate reductase from Rhodobacter capsulatus N22DNAR+ has been purified. It comprises a single type of polypeptide chain with subunit molecular weight 90,000 and does not contain heme. Chlorate is not an alternative substrate. A molybdenum cofactor, of the pterin type found in both nitrate reductases and molybdoenzymes from various sources, is present in nitrate reductase from R. capsulatus at an approximate stoichiometry of 1 molecule per polypeptide chain. This is the first report of the occurrence of the cofactor in a periplasmic enzyme. Trimethylamine-N-oxide reductase activity was fractionated by ion exchange chromatography of periplasmic proteins. The fractionated material was active towards dimethylsulphoxide, chlorate and methionine sulphoxide, but not nitrate. A catalytic polypeptide of molecular weight 46,000 was identified by staining for trimethylamine-N-oxide reductase activity after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. The same polypeptide also stained for dimethylsulphoxide reductase activity which indicates that trimethylamine-N-oxide and dimethylsulphoxide share a common reductase.

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Abbreviations

DMSO:

dimethylsulphoxide

LDS:

lithium dodecyl sulphate

MVH:

reduced methylviologen

PAGE:

polyacrylamide gel electrophoresis

SDS:

sodium dodecyl sulphate

TMAO:

trimethylamine-N-oxide

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Author notes

  1. A. G. McEwan

    Present address: Department of Microbiology, University of Illinois, 61801, Urbana, IL, USA

  2. H. G. Wetzstein

    Present address: Bayer AG, D-5090, Leverkusen, Federal Republic of Germany

  3. O. Meyer

    Present address: Institut für Pflanzenphysiologie und Mikrobiologie, Freie Universität Berlin, Königin-Luise-Str. 12-16a, D-1000, Berlin 33

  4. S. J. Ferguson

    Present address: Department of Biochemistry, University of Oxford, South Parks Road, OX1 3QU, Oxford, UK

Authors and Affiliations

  1. Department of Biochemistry, University of Birmingham, P. O. Box 363, B15 2TT, Birmingham, UK

    A. G. McEwan, H. G. Wetzstein, J. B. Jackson & S. J. Ferguson

  2. Institut für Mikrobiologie der Universität Göttingen, D-3400, Göttingen, Federal Republic of Germany

    O. Meyer

Authors
  1. A. G. McEwan
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  2. H. G. Wetzstein
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  3. O. Meyer
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  4. J. B. Jackson
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  5. S. J. Ferguson
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McEwan, A.G., Wetzstein, H.G., Meyer, O. et al. The periplasmic nitrate reductase of Rhodobacter capsulatus; purification, characterisation and distinction from a single reductase for trimethylamine-N-oxide, dimethylsulphoxide and chlorate. Arch. Microbiol. 147, 340–345 (1987). https://doi.org/10.1007/BF00406130

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  • DOI: https://doi.org/10.1007/BF00406130

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