Pyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeon, Pyrococcus furiosus, functions as a CoA-dependent pyruvate decarboxylase.
Pyruvate ferredoxin oxidoreductase (POR) has been previously purified from the hyperthermophilic archaeon, Pyrococcus furiosus, an organism that grows optimally at 100 degrees C by fermenting carbohydrates and peptides. The enzyme contains thiamine pyrophosphate and catalyzes the oxidative decarboxylation of pyruvate to acetyl-CoA and CO2 and reduces P. furiosus ferredoxin. Here we show that this enzyme also catalyzes the formation of acetaldehyde from pyruvate in a CoA-dependent reaction. Desulfocoenzyme A substituted for CoA showing that the cofactor plays a structural rather than a catalytic role. Ferredoxin was not necessary for the pyruvate decarboxylase activity of POR, nor did it inhibit acetaldehyde production. The apparent Km values for CoA and pyruvate were 0.11 mM and 1.1 mM, respectively, and the optimal temperature for acetaldehyde formation was above 90 degrees C. These data are comparable to those previously determined for the pyruvate oxidation reaction of POR. At 80 degrees C (pH 8.0), the apparent Vm value for pyruvate decarboxylation was about 40% of the apparent Vm value for pyruvate oxidation rate (using P. furiosus ferredoxin as the electron acceptor). Tentative catalytic mechanisms for these two reactions are presented. In addition to POR, three other 2-keto acid ferredoxin oxidoreductases are involved in peptide fermentation by hyperthermophilic archaea. It is proposed that the various aldehydes produced by these oxidoreductases in vivo are used by two aldehyde-utilizing enzymes, alcohol dehydrogenase and aldehyde ferredoxin oxidoreductase, the physiological roles of which were previously unknown.
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Center for Metalloenzyme Studies, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA.
Hutchins
A
A
Sung
S J
SJ
Adams
M W
MW
D000445:Aldehyde Oxidoreductases / Q000378:metabolism; D001105:Archaea / Q000201:enzymology*; D003065:Coenzyme A / Q000378:metabolism; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism; D007658:Ketone Oxidoreductases / Q000378:metabolism*; D050743:Pyruvate Synthase
D005287:Ferredoxin-NADP Reductase; D000445:Aldehyde Oxidoreductases; D007658:Ketone Oxidoreductases; C080010:aldehyde ferredoxin oxidoreductase; D050743:Pyruvate Synthase; D003065:Coenzyme A