Characterization of the nitric oxide reductase from Thermus thermophilus

Lici A. Schurig-Briccio, Padmaja Venkatakrishnan, James Hemp, Carlos Bricio, José Berenguer, Robert B. Gennis

Research output: Contribution to journalArticle

Abstract

Nitrous oxide (N2O) is a powerful greenhouse gas implicated in climate change. The dominant source of atmospheric N2O is incomplete biological dentrification, and the enzymes responsible for the release of N 2O are NO reductases. It was recently reported that ambient emissions of N2O from the Great Boiling Spring in the United States Great Basin are high, and attributed to incomplete denitrification by Thermus thermophilus and related bacterial species [Hedlund BP, et al. (2011) Geobiology 9(6)471-480]. In the present work, we have isolated and characterized the NO reductase (NOR) from T. thermophilus. The enzyme is a member of the cNOR family of enzymes and belongs to a phylogenetic clade that is distinct from previously examined cNORs. Like other characterized cNORs, the T. thermophilus cNOR consists of two subunits, NorB and NorC, and contains a one heme c, one Ca 2+, a low-spin heme b, and an active site consisting of a high-spin heme b and FeB. The roles of conserved residues within the cNOR family were investigated by site-directed mutagenesis. The most important and unexpected result is that the glutamic acid ligand to FeB is not essential for function. The E211A mutant retains 68% of wild-type activity. Mutagenesis data and the pattern of conserved residues suggest that there is probably not a single pathway for proton delivery from the periplasm to the active site that is shared by all cNORs, and that there may be multiple pathways within the T. thermophilus cNOR.

Original languageEnglish (US)
Pages (from-to)12613-12618
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume110
Issue number31
DOIs
StatePublished - Jul 30 2013

Keywords

  • Proton pathway
  • Thermophilic

ASJC Scopus subject areas

  • General

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