title: WOR5, a novel tungsten-containing aldehyde oxidoreductase from Pyrococcus furiosus with a broad substrate Specificity.
abstract: WOR5 is the fifth and last member of the family of tungsten-containing oxidoreductases purified from the hyperthermophilic archaeon Pyrococcus furiosus. It is a homodimeric protein (subunit, 65 kDa) that contains one [4Fe-4S] cluster and one tungstobispterin cofactor per subunit. It has a broad substrate specificity with a high affinity for several substituted and nonsubstituted aliphatic and aromatic aldehydes with various chain lengths. The highest catalytic efficiency of WOR5 is found for the oxidation of hexanal (V(max) = 15.6 U/mg, K(m) = 0.18 mM at 60 degrees C). Hexanal-incubated enzyme exhibits S = 1/2 electron paramagnetic resonance signals from [4Fe-4S]1+ (g values of 2.08, 1.93, and 1.87) and W5+ (g values of 1.977, 1.906, and 1.855). Cyclic voltammetry of ferredoxin and WOR5 on an activated glassy carbon electrode shows a catalytic wave upon addition of hexanal, suggesting that ferredoxin can be a physiological redox partner. The combination of WOR5, formaldehyde oxidoreductase, and aldehyde oxidoreductase forms an efficient catalyst for the oxidation of a broad range of aldehydes in P. furiosus.
authors.affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands.
authors.forename: Loes E
authors.initials: LE
authors.lastname: Bevers
authors.forename: Emile
authors.initials: E
authors.lastname: Bol
authors.forename: Peter-Leon
authors.initials: PL
authors.lastname: Hagedoorn
authors.forename: Wilfred R
authors.initials: WR
authors.lastname: Hagen
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism*; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D002384:Catalysis; D004579:Electron Transport; D008969:Molecular Sequence Data; D020027:Pyrococcus furiosus / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D014414:Tungsten / Q000378:metabolism*
chemical_list: D000447:Aldehydes; D019843:Archaeal Proteins; D000445:Aldehyde Oxidoreductases; D014414:Tungsten