Applied and Environmental Microbiology, November 2001, p. 5122-5126, Vol. 67, No. 11
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.11.5122-5126.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
CEA/Cadarache, DSV, DEVM, Laboratoire de Bioénergétique Cellulaire, 13108 St. Paul lez Durance Cedex,1 and Laboratoire de Cristallographie et Cristallogénèse des Protéines, Institut de Biologie Structurale JP Ebel CEA-CNRS, 38027 Grenoble Cedex 1,2 France
Received 29 March 2001/Accepted 9 August 2001
Preliminary studies showed that the periplasmic nitrate reductase
(Nap) of Rhodobacter sphaeroides and the membrane-bound nitrate reductases of Escherichia coli are able to reduce
selenate and tellurite in vitro with benzyl viologen as an electron
donor. In the present study, we found that this is a general feature of
denitrifiers. Both the periplasmic and membrane-bound nitrate reductases of Ralstonia eutropha, Paracoccus denitrificans,
and Paracoccus pantotrophus can utilize potassium selenate
and potassium tellurite as electron acceptors. In order to characterize
these reactions, the periplasmic nitrate reductase of R. sphaeroides f. sp. denitrificans IL106 was histidine
tagged and purified. The Vmax and
Km were determined for nitrate, tellurite, and
selenate. For nitrate, values of 39 µmol · min
1 · mg
1 and 0.12 mM were obtained
for Vmax and Km,
respectively, whereas the Vmax values for
tellurite and selenate were 40- and 140-fold lower, respectively. These
low activities can explain the observation that depletion of the
nitrate reductase in R. sphaeroides does not modify the MIC
of tellurite for this organism.
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