Purification and some properties of an acryloyl-CoA reductase of Clostridium kluyveri.
Acryloyl-CoA reductase, a presumably previously unknown soluble enzyme, is present in Clostridium kluyveri. It catalyses the reduction of the carbon-carbon double bond of acryloyl-CoA or ethyl vinyl ketone and other alpha, beta-unsaturated carbonyl compounds at the expense of reduced methylviologen. On the basis of a Vmax/Km ratio, which is at least 18 times higher than that for the next best substrate (E)-2-butenoyl-CoA, the enzyme is called acryloyl-CoA reductase. A purity of over 90% was achieved. The apparent molecular mass, as determined by gel chromatography, is 28.4 kDa. Dodecyl sulfate gel electrophoresis shows subunits with a molecular mass of 14.2 kDa. Based on a molecular mass of 28.4 kDa about 1.5 mol FMN have been observed. Less than 0.2 g-atom iron per mol protein were determined. Ferredoxin or flavodoxin seem to be able to carry electrons from hydrogenase to the acryloyl-CoA reductase. The addition of hydrogen to the alpha-carbon of ethyl vinyl ketone occurs from the re-side.
D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D003013:Clostridium / Q000201:enzymology*; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D010088:Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity; D013696:Temperature
D010088:Oxidoreductases; C047341:acryloyl-CoA reductase