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Volume 166, Issue 10

is essential for whole-cell NO reduction in but not for assembly of copper centres of nitrous oxide reductase Open Access

Abstract

Nitrous oxide (NO) is a potent greenhouse gas that is produced naturally as an intermediate during the process of denitrification carried out by some soil bacteria. It is consumed by nitrous oxide reductase (NOR), the terminal enzyme of the denitrification pathway, which catalyses a reduction reaction to generate dinitrogen. NOR contains two important copper cofactors (Cu and Cu centres) that are essential for activity, and in copper-limited environments, NOR fails to function, contributing to rising levels of atmospheric NO and a major environmental challenge. Here we report studies of , one of eight genes in the cluster of the soil dwelling α-proteobaterium . A Δ deletion mutant failed to reduce NO under both copper-sufficient and copper-limited conditions, demonstrating that NosX plays an essential role in NOR activity. NOR isolated from -deficient cells was found to be unaffected in terms of the assembly of its copper cofactors, and to be active in assays, indicating that NosX is not required for the maturation of the enzyme; in particular, it plays no part in the assembly of either of the Cu and Cu centres. Furthermore, quantitative Reverse Transcription PCR (qRT-PCR) studies showed that NosX does not significantly affect the expression of the NOR-encoding gene. NosX is a homologue of the FAD-binding protein ApbE from , which functions in the flavinylation of another NOR accessory protein, NosR. Thus, it is likely that NosX is a system-specific maturation factor of NosR, and so is indirectly involved in maintaining the reaction cycle of NOR and cellular NO reduction.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): copper , denitrification , nitrous oxide and nitrous oxide reductase
Funding
This study was supported by the:
  • Biotechnology and Biological Sciences Research Council (Award NRP DTP)
    • Principle Award Recipient: Sophie P Bennett
  • EU (Award 316472)
    • Principle Award Recipient: Manuel J Soriano-Laguna
  • Biotechnology and Biological Sciences Research Council (Award BB/S008942/1)
    • Principle Award Recipient: Andrew J Gates
  • Biotechnology and Biological Sciences Research Council (Award BB/M00256X/1)
    • Principle Award Recipient: Andrew J Gates
  • Biotechnology and Biological Sciences Research Council (Award BB/L022796/1)
    • Principle Award Recipient: David J Richardson
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2020-09-04
2024-04-19
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nosX is essential for whole-cell N2O reduction in Paracoccus denitrificans but not for assembly of copper centres of nitrous oxide reductase
Microbiology 166, 909 (2020); https://doi.org/10.1099/mic.0.000955
/content/journal/micro/10.1099/mic.0.000955
/content/journal/micro/10.1099/mic.0.000955
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