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Lookup NU author(s): Antonius Pierik, Emeritus Professor Bernard Golding
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Acryloyl-CoA reductase from Clostridium propionicum catalyses the irreversible NADH-dependent formation of propionyl-CoA from acryloyl-CoA. Purification yielded a heterohexadecameric yellow-greenish enzyme complex [(α2βγ)4; molecular mass 600 ± 50 kDa] composed of a propionyl-CoA dehydrogenase (α2, 2 × 40 kDa) and an electron-transferring flavoprotein (ETF; β, 38 kDa; γ, 29 kDa). A flavin content (90% FAD and 10% FMN) of 2.4 mol per α2βγ subcomplex (149 kDa) was determined. A substrate alternative to acryloyl-CoA (Km = 2 ± 1 μM; kcat = 4.5 s-1 at 100 μM NADH) is 3-buten-2-one (methyl vinyl ketone; Km = 1800 μM kcat 29 s-1 at 300 μM NADH). The enzyme complex exhibits acyl-CoA dehydrogenase activity with propionyl-CoA (Km = 50 μM; kcat = 2.0 s-1) or butyryl-CoA (Km = 100 μM; kcat = 3.5 s-1) as electron donor and 200 μM ferricenium hexafluorophosphate as acceptor. The enzyme also catalysed the oxidation of NADH by iodonitrosotetrazolium chloride (diaphorase activity) or by air, which led to the formation of H2O2 (NADH oxidase activity). The N-terminus of the dimeric propionyl-CoA dehydrogenase subunit is similar to those of butyryl-CoA dehydrogenases from several clostridia and related anaerobes (up to 55% sequence identity). The N-termini of the β and γ subunits share 40% and 35% sequence identities with those of the A and B subunits of the ETF from Megasphaera elsdenii, respectively, and up to 60% with those of putative ETFs from other anaerobes. Acryloyl-CoA reductase from C. propionicum has been characterized as a soluble enzyme, with kinetic properties perfectly adapted to the requirements of the organism. The enzyme appears not to be involved in anaerobic respiration with NADH or reduced ferredoxin as electron donors. There is no relationship to the trans-2-enoyl-CoA reductases from various organisms or the recently described acryloyl-CoA reductase activity of propionyl-CoA synthase from Chloroflexus aurantiacus.
Author(s): Hetzel M, Brock M, Selmer T, Pierik AJ, Golding BT, Buckel W
Publication type: Article
Publication status: Published
Journal: European Journal of Biochemistry
Year: 2003
Volume: 270
Issue: 5
Pages: 902-910
Print publication date: 01/03/2003
ISSN (print): 0014-2956
ISSN (electronic): 1742-4658
Publisher: Wiley-Blackwell Publishing
URL: http://dx.doi.org/10.1046/j.1432-1033.2003.03450.x
DOI: 10.1046/j.1432-1033.2003.03450.x
PubMed id: 12603323
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