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Biogenesis of nitrous oxide reductase – a key enzyme in controlling climate change (LEBRUNNU19SF)


Project Description

Atmospheric nitrous oxide (N2O) not only destroys stratospheric ozone, it also accounts for ~10% of the impact due to greenhouse gases. Much of the N2O generated arises from activity of soil bacteria that are able to grow anaerobically through denitrification, a process central to the biogeochemical cycling of nitrogen, in which nitrate is reduced stepwise to dinitrogen (N2) via nitrite, nitric oxide (NO), and N2O.  The last step of the pathway, reduction of N2O to N2, is catalysed by the enzyme nitrous oxide reductase (N2OR); failure of this step to occur is the major cause of N2O release into the atmosphere.  Increased levels of denitrification and hence N2O emissions have arisen through the increased use in agriculture of nitrogenous fertilisers. 
 
N2OR contains two distinct copper cofactors: CuA, is a thiol-bridged dinuclear copper centre, and CuZ, a unique copper-sulfur cluster.  Biogenesis of N2OR is dependent on a number of assembly proteins whose functions are not well defined.  The aim of this PhD project is to provide new molecular detail of the trafficking of copper and sulfide for incorporation into CuA and CuZ centres and will involve in vitro biophysical studies of proteins from the model denitrifier Paracoccus denitrificans that are known to function directly in N2OR assembly, or in copper/sulfur trafficking in general.  The work will lead to new insights into the biogenesis of N2OR, feeding into efforts towards maintaining and maximising N2OR activity in soils, with the long term benefit of reducing N2O emissions.  This multi-disciplinary project will be based in the Le Brun Lab at UEA and will involve protein purification, biochemistry, native mass spectrometry, spectroscopy and kinetics, offering excellent training potential for the appointed student within a supportive and stimulating environment. For informal enquiries, please contact Prof Nick Le Brun ().

Applications are processed as soon as they are received and the project may be filled before the closing date, so early application is encouraged.

Project Start Date: Oct 2019
Mode of Study: Full-time
Acceptable First Degree: Chemistry, Biochemistry or related subjects
Minimum Entry Requirements: UK 2:1

Funding Notes

This PhD project is offered on a self-funding basis. It is open to applicants with funding or those applying to funding sources.  Details of tuition fees can be found at View Website.  
 
A bench fee is also payable on top of the tuition fee to cover specialist equipment or laboratory costs required for the research.  The amount charged annually will vary considerably depending on the nature of the project and applicants should contact the primary supervisor for further information about the fee associated with the project. 

References

i) Thomson et al (2012) Phil Trans Royal Soc B 367, 1157-1168 
ii) Rasmussen et al (2000) Biochemistry 39, 12753-12756 
iii) Crack et al (2017) Proc. Natl. Acad. Sci. U.S.A. 114, E3215-E3223 
iv) Kay et al (2017) Chem. Comm. 53, 1397-1400 
v) Le Brun, N. E. (2013) Binding, Transport and Storage of Copper in Prokaryotes, in ‘Binding, Transport and Storage of Metal Ions in Biological Cells’, Maret, W. and Wedd, A. G. Eds. RSC Publishing, pp 461 - 499

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