Denitrification is a distinct means of energy conservation, making use of N oxides as terminal electron acceptors for cellular bioenergetics under anaerobic, microaerophilic, and occasionally aerobic conditions. The process is an essential branch of the global N cycle, reversing dinitrogen fixation, and is associated with chemolithotrophic, phototrophic, diazotrophic, or organotrophic metabolism but generally not with obligately anaerobic life. Discovered more than a century ago and believed to be exclusively a bacterial trait, denitrification has now been found in halophilic and hyperthermophilic archaea and in the mitochondria of fungi, raising evolutionarily intriguing vistas. Important advances in the biochemical characterization of denitrification and the underlying genetics have been achieved with Pseudomonas stutzeri, Pseudomonas aeruginosa, Paracoccus denitrificans, Ralstonia eutropha, and Rhodobacter sphaeroides. Pseudomonads represent one of the largest assemblies of the denitrifying bacteria within a single genus, favoring their use as model organisms. Around 50 genes are required within a single bacterium to encode the core structures of the denitrification apparatus. Much of the denitrification process of gram-negative bacteria has been found confined to the periplasm, whereas the topology and enzymology of the gram-positive bacteria are less well established. The activation and enzymatic transformation of N oxides is based on the redox chemistry of Fe, Cu, and Mo. Biochemical breakthroughs have included the X-ray structures of the two types of respiratory nitrite reductases and the isolation of the novel enzymes nitric oxide reductase and nitrous oxide reductase, as well as their structural characterization by indirect spectroscopic means. This revealed unexpected relationships among denitrification enzymes and respiratory oxygen reductases. Denitrification is intimately related to fundamental cellular processes that include primary and secondary transport, protein translocation, cytochrome c biogenesis, anaerobic gene regulation, metalloprotein assembly, and the biosynthesis of the cofactors molybdopterin and heme D1. An important class of regulators for the anaerobic expression of the denitrification apparatus are transcription factors of the greater FNR family. Nitrate and nitric oxide, in addition to being respiratory substrates, have been identified as signaling molecules for the induction of distinct N oxide-metabolizing enzymes.

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Cell biology and molecular basis of denitrification.

Mechanistic aspects of the reactions of nitric oxide with transition-metal complexes.

Department of Chemistry, The Johns Hopkins University, Charles and 34th Streets, Baltimore, Maryland 21218, Kluyver Laboratory for Biotechnology, Delft University of Technology, 2628 BC Delft, The Netherlands, Department of Biochemistry and Molecular Biology, OGI School of Science and Engineering at OHSU, Beaverton, Oregon 97006, and Department of Microbiology and Molecular Genetics, University of California, Los Angeles, California 90095

Nitric Oxide in Biological Denitrification: Fe/Cu Metalloenzyme and Metal Complex NOx Redox Chemistry

The chemical reduction of nitrate in aqueous solution

The Denitrifying Prokaryotes

In an effort to provide precedence for postulated intermediates in copper-protein-mediated nitrite reduction, a series of novel complexes containing the CuI−NO2- unit, including monocopper(I), dicopper(I,I), and mixed valence dicopper(I,II) and copper(I)−zinc(II) species, were prepared, fully characterized, and subjected to reactivity studies designed to probe their ability to produce nitric oxide. Treatment of solutions of [LCu(CH3CN)]PF6 (L = Li-Pr3, 1,4,7-triisopropyl-1,4,7-triazacyclononane, or LBn3, 1,4,7-tribenzyl-1,4,7-triazacyclononane) in MeOH with excess NaNO2 yielded the novel dicopper(I,I) complexes [(LCu)2(μ-NO2)]PF6. The complex with L = Li-Pr3 was cleaved by PPh3 to afford [Li-Pr3Cu(PPh3)]PF6 and Li-Pr3Cu(NO2), a structural model for the substrate adduct of copper nitrite reductase. Oxidation of the dicopper(I,I) compound (L = Li-Pr3) with (Cp2Fe)(PF6) in CH2Cl2 yielded the deep red, mixed valent, dicopper(I,II) species [(Li-Pr3Cu)2(μ-NO2)](PF6)2, which was structurally characterized as its...

Synthetic Modeling of Nitrite Binding and Activation by Reduced Copper Proteins. Characterization of Copper(I)−Nitrite Complexes That Evolve Nitric Oxide

Alan J. Gengenbach

Papers

Synthetic Modeling of Nitrite Binding and Activation by Reduced Copper Proteins. Characterization of Copper(I)−Nitrite Complexes That Evolve Nitric Oxide