Extremophiles

, Volume 13, Issue 4, pp 687–693 | Cite as

Characterization of a soluble oxidoreductase from the thermophilic bacterium Carboxydothermus ferrireducens

Original Paper
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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.

Keywords

Anaerobic bacteria Biochemical characterization Thermophiles and thermophilic enzymes Oxidoreductase Metal reduction Oxidative stress Gram-type positive Flavoprotein Quinones 

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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Notes

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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Copyright information

© Springer 2009

Authors and Affiliations

  • Rob Uche Onyenwoke
    • 1
    • 3
  • R. Geyer
    • 2
  • Juergen Wiegel
    • 1
  1. 1.Department of MicrobiologyThe University of GeorgiaAthensUSA
  2. 2.LC-MS Support, Core Pharma-Central Europe, Applera Europe B.V., Rotkreuz BranchRotkreuzSwitzerland
  3. 3.Neuroscience Center, UNC School of MedicineUniversity of North CarolinaChapel HillUSA

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