WOR5, a novel tungsten-containing aldehyde oxidoreductase from Pyrococcus furiosus with a broad substrate specificity

被引:41
作者
Bevers, LE [1 ]
Bol, E [1 ]
Hagedoorn, PL [1 ]
Hagen, WR [1 ]
机构
[1] Delft Univ Technol, Dept Biotechnol, NL-2628 BC Delft, Netherlands
关键词
D O I
10.1128/JB.187.20.7056-7061.2005
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
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页码:7056 / 7061
页数:6
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