Nitrate ammonification by Nautilia profundicola AmH: experimental evidence consistent with a free hydroxylamine intermediate
- 1School of Marine Science and Policy, University of Delaware, Newark, DE, USA
- 2Delaware Biotechnology Institute, University of Delaware, Newark, DE, USA
- 3Department of Biological Sciences, Clemson University, Clemson, SC, USA
- 4Department of Biology, University of North Carolina, Charlotte, NC, USA
The process of nitrate reduction via nitrite controls the fate and bioavailability of mineral nitrogen within ecosystems; i.e., whether it is retained as ammonium (ammonification) or lost as nitrous oxide or dinitrogen (denitrification). Here, we present experimental evidence for a novel pathway of microbial nitrate reduction, the reverse hydroxylamine:ubiquinone reductase module (reverse-HURM) pathway. Instead of a classical ammonia-forming nitrite reductase that performs a 6 electron-transfer process, the pathway is thought to employ two catalytic redox modules operating in sequence: the reverse-HURM reducing nitrite to hydroxylamine followed by a hydroxylamine reductase that converts hydroxylamine to ammonium. Experiments were performed on Nautilia profundicola strain AmH, whose genome sequence led to the reverse-HURM pathway proposal. N. profundicola produced ammonium from nitrate, which was assimilated into biomass. Furthermore, genes encoding the catalysts of the reverse-HURM pathway were preferentially expressed during growth of N. profundicola on nitrate as an electron acceptor relative to cultures grown on polysulfide as an electron acceptor. Finally, nitrate-grown cells of N. profundicola were able to rapidly and stoichiometrically convert high concentrations of hydroxylamine to ammonium in resting cell assays. These experiments are consistent with the reverse-HURM pathway and a free hydroxylamine intermediate, but could not definitively exclude direct nitrite reduction to ammonium by the reverse-HURM with hydroxylamine as an off-pathway product. N. profundicola and related organisms are models for a new pathway of nitrate ammonification that may have global impact due to the wide distribution of these organisms in hypoxic environments and symbiotic or pathogenic associations with animal hosts.
Keywords: hydroxylamine, hydroxylamine oxidoreductase, nitrite, Epsilonproteobacteria, nitrate ammonification
Citation: Hanson TE, Campbell BJ, Kalis KM, Campbell MA and Klotz MG (2013) Nitrate ammonification by Nautilia profundicola AmH: experimental evidence consistent with a free hydroxylamine intermediate. Front. Microbiol. 4:180. doi: 10.3389/fmicb.2013.00180
Received: 02 May 2013; Accepted: 15 June 2013;
Published online: 04 July 2013.
Copyright © 2013 Hanson, Campbell, Kalis, Campbell and Klotz. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc.
*Correspondence: Barbara J. Campbell, Department of Biological Sciences, Clemson University, 132 Long Hall, Clemson, SC 29634, USA e-mail: firstname.lastname@example.org