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Elucidation of reaction mechanisms in the anaerobic degradation of hydrocarbons

Subject Area Metabolism, Biochemistry and Genetics of Microorganisms
Term from 2009 to 2016
Project identifier Deutsche Forschungsgemeinschaft (DFG) - Project number 71718654
 
The first part of the proposal aims to deepen our understanding of anaerobic alkane functionalization by the use of substrate analogues in enzymatic studies supported by model studies. The synthetic team at Newcastle will provide labeled substrates (e.g. deuterated decanes) and substrate analogues (e.g. dialkylcyclopropanes) to AG Rabus and AG Wilkes for enzymatic experiments. This work will extend the highly successful cooperation achieved in the first phase of the research. Intermediates for enzymatic studies will also be provided to the Marburg team (substituted malonic and succinic acids for conversion into coenzyme A esters) and AG Heider (‘chiral’ toluene). A new direction will be the development of experimental and theoretical models to improve understanding of the mechanisms of alkane activation. The Newcastle group will continue to provide support to other groups within this SSP (e.g. AG Boll and AG Meckenstock) by synthesizing key intermediates and products, and by giving advice on mechanistic issues in the context of novel catalytic pathways (e.g. benzene carboxylation). For the second part of the proposal the Marburg team will investigate the metabolism of Syntrophus aciditrophicus. The question as to how benzoate degradation and synthesis can proceed via the same enzymes will be studied using the glutaryl-CoA dehydrogenase/electron transferring flavoprotein complex (Gdh/Etf) and the Na+ pumping glutaconyl-CoA decarboxylase (Gcd) as examples. The Marburg team have cloned and functionally expressed four out of six genes coding for these enzymes. The planned studies should reveal whether reduced ferredoxin can drive the oxidation of glutaryl-CoA by NAD+ or whether menaquinone acts as electron acceptor. For reversal of the decarboxylation the source of ΔμNa+ will be identified. The substrate specificity of (Gdh/Etf) will indicate whether the enzyme can be applied for a bio-based production of adipic acid. Finally, the pathway of glutamate synthesis via Re-citrate synthase will be analyzed in more detail.
DFG Programme Priority Programmes
International Connection United Kingdom
 
 

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