The catalytic cycle of nitrous oxide reductase — The enzyme that catalyzes the last step of denitrification

Cíntia Carreira, Sofia R. Pauleta, Isabel Moura

Research output: Contribution to journalShort surveypeer-review

30 Citations (Scopus)


The reduction of the potent greenhouse gas nitrous oxide requires a catalyst to overcome the large activation energy barrier of this reaction. Its biological decomposition to the inert dinitrogen can be accomplished by denitrifiers through nitrous oxide reductase, the enzyme that catalyzes the last step of the denitrification, a pathway of the biogeochemical nitrogen cycle. Nitrous oxide reductase is a multicopper enzyme containing a mixed valence CuA center that can accept electrons from small electron shuttle proteins, triggering electron flow to the catalytic sulfide-bridged tetranuclear copper “CuZ center”. This enzyme has been isolated with its catalytic center in two forms, CuZ*(4Cu1S) and CuZ(4Cu2S), proven to be spectroscopic and structurally different. In the last decades, it has been a challenge to characterize the properties of this complex enzyme, due to the different oxidation states observed for each of its centers and the heterogeneity of its preparations. The substrate binding site in those two “CuZ center” forms and which is the active form of the enzyme is still a matter of debate. However, in the last years the application of different spectroscopies, together with theoretical calculations have been useful in answering these questions and in identifying intermediate species of the catalytic cycle. An overview of the spectroscopic, kinetics and structural properties of the two forms of the catalytic “CuZ center” is given here, together with the current knowledge on nitrous oxide reduction mechanism by nitrous oxide reductase and its intermediate species.

Original languageEnglish
Pages (from-to)423-434
Number of pages12
JournalJournal of Inorganic Biochemistry
Publication statusPublished - Dec 2017


  • Catalytic cycle
  • Copper enzyme
  • CuZ
  • Denitrification
  • Greenhouse effect
  • Nitrous oxide reductase


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