Nitric Oxide Production from Nitrite Reduction and Hydroxylamine Oxidation by Copper-containing Dissimilatory Nitrite Reductase (NirK) from the Aerobic Ammonia-oxidizing Archaeon, Nitrososphaera viennensis

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Aerobic ammonia-oxidizing archaea (AOA) play a crucial role in the global nitrogen cycle by oxidizing ammonia to nitrite, and nitric oxide (NO) is a key intermediate in AOA for sustaining aerobic ammonia oxidation activity. We herein heterologously expressed the NO-forming, copper-containing, dissimilatory nitrite reductase (NirK) from Nitrososphaera viennensis and investigated its enzymatic properties. The recombinant protein catalyzed the reduction of ¹⁵NO₂⁻ to ¹⁵NO, the oxidation of hydroxylamine (¹⁵NH₂OH) to ¹⁵NO, and the production of ^14–15N₂O from ¹⁵NH₂OH and ¹⁴NO₂⁻. To the best of our knowledge, the present study is the first to document the enzymatic properties of AOA NirK.