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Characterization of a soluble oxidoreductase from the thermophilic bacterium Carboxydothermus ferrireducens

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Abstract

An NAD(P)H-dependent oxidoreductase has been purified approximately 40-fold from the soluble protein fraction of the dissimilatory iron-reducing, anaerobic, thermophilic bacterium Carboxydothermus ferrireducens. The enzyme, a flavoprotein, has broad-substrate specificity—reducing Fe3+, Cr6+, and AQDS with rates of 0.31, 0.33, and 3.3 U mg−1 protein and calculated NADH oxidation turnover numbers of 0.25, 0.25, and 2.5 s−1, respectively. Numerous quinones are reduced via a two-electron transfer from NAD(P)H to quinone, thus participating in managing oxidative stress by avoiding the formation of semiquinone radicals.

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Abbreviations

NQO1:

NAD(P)H:quinone oxidoreductase 1

CFOR:

Carboxydothermus ferrireducens NAD(P)H-dependent oxidoreductase

AQDS:

9,10-Anthraquinone-2,6-disulfonate

MES:

2-(N-morpholino)ethanesulfonic acid

MOPS:

3-(N-morpholino)propanesulfonic acid

TAPS:

N-Tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid

QH2 :

Reduced quinones

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Acknowledgments

We thank Drs. David Siegel and J. Samuel Zigler for the generous gifts of recombinant human NAD(P)H:quinone oxidoreductase and guinea pig zeta-crystallin protein, respectively, and Harry Dailey, Michael W.W. Adams, William B. Whitman, and Robert J. Maier for helpful discussions.

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Author notes

  1. Rob Uche Onyenwoke

    Present address: Neuroscience Center, UNC School of Medicine, University of North Carolina, Chapel Hill, NC, USA

Authors and Affiliations

  1. Department of Microbiology, The University of Georgia, 212 Biological Sciences Bldg, 1000 Cedar Street, Athens, GA, 30602, USA

    Rob Uche Onyenwoke & Juergen Wiegel

  2. LC-MS Support, Core Pharma-Central Europe, Applera Europe B.V., Rotkreuz Branch, Grundstrasse 10, 6343, Rotkreuz, Switzerland

    R. Geyer

Authors
  1. Rob Uche Onyenwoke
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  2. R. Geyer
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  3. Juergen Wiegel
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Corresponding author

Correspondence to Juergen Wiegel.

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Communicated by A. Driessen.

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Onyenwoke, R.U., Geyer, R. & Wiegel, J. Characterization of a soluble oxidoreductase from the thermophilic bacterium Carboxydothermus ferrireducens . Extremophiles 13, 687–693 (2009). https://doi.org/10.1007/s00792-009-0255-1

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  • DOI: https://doi.org/10.1007/s00792-009-0255-1

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