title: (2S)-Methylsuccinyl-CoA dehydrogenase closes the ethylmalonyl-CoA pathway for acetyl-CoA assimilation.
abstract: Many organic substrates are metabolized via acetyl-coenzyme A (CoA) and enter central carbon metabolism at the level of this compound. We recently described the outlines of the ethylmalonyl-CoA pathway, a new acetyl-CoA assimilation strategy that operates in a number of bacteria such as Rhodobacter sphaeroides, Methylobacterium extorquens and streptomycetes and replaces the glyoxylate cycle. This new pathway involves a unique central reaction sequence catalysed by characteristic enzymes. Here, we identified and characterized (2S)-methylsuccinyl-CoA dehydrogenase from R. sphaeroides, a flavin adenine dinucleotide-containing enzyme that catalyses the last unknown step in the central part of the ethylmalonyl-CoA pathway, the oxidation of (2S)-methylsuccinyl-CoA to mesaconyl-(C1)-CoA. This enzyme is highly specific for its substrate and forms a distinct subgroup within the superfamily of flavin-dependent acyl-CoA dehydrogenases. Homology modelling and comparative sequence analyses with well-studied members of this superfamily identified amino acids that may contribute to the narrow substrate specificity of (2S)-methylsuccinyl-CoA dehydrogenase. The central part of the ethylmalonyl-CoA pathway was reconstituted in vitro using four recombinant enzymes. By this work, the ethylmalonyl-CoA pathway and its stereochemical course have been completely solved. This allowed defining the minimum set of enzymes necessary for its operation and to screen for further organisms following this acetyl-CoA assimilation strategy.
authors.affiliation: Mikrobiologie, Institut für Biologie II, Schänzlestr 1, Albert-Ludwigs-Universität Freiburg, Freiburg i Br, Germany.
authors.forename: Tobias J
authors.initials: TJ
authors.lastname: Erb
authors.forename: Georg
authors.initials: G
authors.lastname: Fuchs
authors.forename: Birgit E
authors.initials: BE
authors.lastname: Alber
mesh_terms: D000105:Acetyl Coenzyme A / Q000378:metabolism*; D000214:Acyl Coenzyme A / Q000378:metabolism*; D000595:Amino Acid Sequence; D003067:Coenzymes / Q000494:pharmacology; D005182:Flavin-Adenine Dinucleotide / Q000494:pharmacology; D053858:Metabolic Networks and Pathways; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000378:metabolism*; D012242:Rhodobacter sphaeroides / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D000214:Acyl Coenzyme A; D003067:Coenzymes; C102829:ethylmalonyl-coenzyme A; C102828:methylsuccinyl-coenzyme A; D005182:Flavin-Adenine Dinucleotide; D000105:Acetyl Coenzyme A; D010088:Oxidoreductases