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The Ammonia Transport, Retention and Futile Cycling Problem in Cyanobacteria

  • Physiology and Biotechnology
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Abstract

Ammonia is the preferred nitrogen source for many algae including the cyanobacterium Synechococcus elongatis (Synechococcus R-2; PCC 7942). Modelling ammonia uptake by cells is not straightforward because it exists in solution as NH3 and NH +4 . NH3 is readily diffusible not only via the lipid bilayer but also through aquaporins and other more specific porins. On the other hand, NH +4 requires cationic transporters to cross a membrane. Significant intracellular ammonia pools (≈1–10 mol m−3) are essential for the synthesis of amino acids from ammonia. The most common model envisaged for how cells take up ammonia and use it as a nitrogen source is the “pump–leak model” where uptake occurs through a simple diffusion of NH3 or through an energy-driven NH +4 pump balancing a leak of NH3 out of the cell. The flaw in such models is that cells maintain intracellular pools of ammonia much higher than predicted by such models. With caution, [14C]-methylamine can be used as an analogue tracer for ammonia and has been used to test various models of ammonia transport and metabolism. In this study, simple “proton trapping” accumulation by the diffusion of uncharged CH3NH2 has been compared to systems where CH3NH +3 is taken up through channels, driven by the membrane potential (ΔU i,o) or the electrochemical potential for Na+ (ΔμNa +i,o ). No model can be reconciled with experimental data unless the permeability of CH3NH2 across the cell membrane is asymmetric: permeability into the cell is very high through gated porins, whereas permeability out of the cell is very low (≈40 nm s−1) and independent of the extracellular pH. The best model is a Na +in /CH3NH +3 in co-porter driven by ΔμNa +i,o balancing synthesis of methylglutamine and a slow leak governed by Ficks law, and so there is significant futile cycling of methylamine across the cell membrane to maintain intracellular methylamine pools high enough for fixation by glutamine synthetase. The modified pump–leak model with asymmetric permeability of the uncharged form is a viable model for understanding ammonia uptake and retention in plants, free-living microbes and organisms in symbiotic relationships.

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Abbreviations

A:

An amine of general formula R–NH2 in the uncharged form

BIS-TRIS-PROPANE:

1,3-Bis[tris(hydroxymethyl)methylamino]propane

CAPS:

3-(Cyclohexylamino)-1-propanesulfonic acid

HA+ :

An amine (R-NH2) in the charged form R–NH +3

MES:

4-Morpholineethanesulphonic acid

PA:

Permeability of an uncharged amine (R–NH2)

PHA+ :

Permeability of an amine cation (R–NH +3 )

PNH3 :

Permeability of NH3 across cell membrane at a specified pH

PMA:

Permeability of CH3NH2 across cell membrane at a specified pH

PNH +4 :

Goldman permeability of NH +4 across the cell membrane

PMA+ :

Goldman permeability of CH3NH +3 across the cell membrane

PPFD:

Photosynthetic photon fluence density (400–700 nm)

i (subscript):

Refers to the inside of the cells

o (subscript):

Refers to the outside of the cells or bulk electrolyte

ΔU i,o :

The membrane potential gradient inside vs. outside

ΔμNa +i,o :

The sodium motive force or sodium electrochemical potential

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Acknowledgments

The author wishes to thank Profs. IR Kennedy and AWD Larkum (University of Sydney, Australia) for their interest and encouragement in this project. Dr. Michael Merrick (Department of Molecular Microbiology, John Innes Centre, Norwich University) provided very useful up-to-date information on aquaporins.

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  1. Faculty of Technology & Environment, Prince of Songkla University—Phuket Campus, Kathu, Phuket, 83120, Thailand

    Raymond J. Ritchie

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Ritchie, R.J. The Ammonia Transport, Retention and Futile Cycling Problem in Cyanobacteria. Microb Ecol 65, 180–196 (2013). https://doi.org/10.1007/s00248-012-0111-1

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