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Literature summary extracted from

  • Mukund, S.; Adams, M.W.W.
    The novel tungsten-iron-sulfur protein of the hyperthermophilic archaebacterium, Pyrococcus furiosus, is an aldehyde ferredoxin oxidoreductase. Evidence for its participation in a unique glycolytic pathway (1991), J. Biol. Chem., 266, 14208-14216.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.2.7.5 additional information no effect of CoA Pyrococcus furiosus

General Stability

EC Number General Stability Organism
1.2.7.5 10% glycerol and 2 mM DTT stabilize during purification Pyrococcus furiosus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.2.7.5 arsenite 50% inhibition at 1 mM Pyrococcus furiosus
1.2.7.5 crotonaldehyde substrate inhibition above 0.2 mM Pyrococcus furiosus
1.2.7.5 cyanide 50% inhibition at 8 mM Pyrococcus furiosus
1.2.7.5 iodoacetate 50% inhibition at 0.2 mM Pyrococcus furiosus
1.2.7.5 additional information no effect of CoA Pyrococcus furiosus
1.2.99.6 crotonaldehyde above 0.2 mM Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.7.5 0.04
-
 crotonaldehyde pH 8.4, 80°C, with methyl viologen as electron acceptor Pyrococcus furiosus
1.2.7.5 1
-
 glyceraldehyde pH 8.4, 65°C, with methyl viologen as electron acceptor Pyrococcus furiosus
1.2.99.6 0.04
-
 crotonaldehyde
-
 Pyrococcus furiosus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.2.7.5 Fe2+ 6.6 gatoms per 85000g of protein Pyrococcus furiosus
1.2.7.5 Molybdenum part of a tungsten-molybdopterin cofactor Pyrococcus furiosus
1.2.7.5 Tungsten part of a tungsten-molybdopterin cofactor Pyrococcus furiosus
1.2.7.5 Tungsten 1.0 gatoms per 85000g of protein Pyrococcus furiosus
1.2.99.6 Iron contains 7 gatom of Fe per mol of enzyme Pyrococcus furiosus
1.2.99.6 Tungsten contains 1 gatom of tungsten per mol of enzyme Pyrococcus furiosus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.2.7.5 80000
-
 1 * 80000, SDS-PAGE Pyrococcus furiosus
1.2.7.5 90000
-
 gel filtration Pyrococcus furiosus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.7.5 an aldehyde + H2O + oxidized ferredoxin Pyrococcus furiosus
-
 an acid + H+ + reduced ferredoxin
-
 ?
1.2.7.5 glyceraldehyde + H2O + oxidized ferredoxin Pyrococcus furiosus involved in glycolysis glycerate + H+ + reduced ferredoxin
-
 ir
1.2.99.6 additional information Pyrococcus furiosus its physiological role is proposed as the oxidation of glyceraldehyde to glycerate in a unique, partially nonphosphorylated glycolytic pathway that generates acetyl-CoA from glucose without participation of nicotinamide nucleotides ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.2.7.5 Pyrococcus furiosus
-
 hyperthermophilic archaeon
-
1.2.7.5 Pyrococcus furiosus
-
 strain DSM 3638
-
1.2.99.6 Pyrococcus furiosus
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.2.7.5 sensitive against O2, dithionite is added to avoid trace contamination with O2 during purification and enzyme assay Pyrococcus furiosus
1.2.99.6 90% loss of activity after 10 min exposure to air Pyrococcus furiosus

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.7.5 to homogeneity under strict anaerobic conditions Pyrococcus furiosus
1.2.99.6
-
 Pyrococcus furiosus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.2.7.5 53.6
-
 purified enzyme, substrate crotonaldehyde with methyl viologen as electron acceptor Pyrococcus furiosus
1.2.99.6 53.6
-
-
 Pyrococcus furiosus

Storage Stability

EC Number Storage Stability Organism
1.2.7.5 -196°C, pure enzyme pellet is thawed anaerobically after storage in liquid N2, no loss of activity for several months Pyrococcus furiosus
1.2.7.5 4°c or 23°C, pure enzyme, 50 mM Tris-HCl, pH 8.0, 2 mM sodium dithionite, 2 mM DTT, 10% v/v glycerol, loss of 25% activity after 6 h under strict anaerobic conditions Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.7.5 acetaldehyde + H2O + oxidized ferredoxin
-
 Pyrococcus furiosus acetate + H+ + reduced ferredoxin
-
 ir
1.2.7.5 an aldehyde + H2O + oxidized ferredoxin
-
 Pyrococcus furiosus an acid + H+ + reduced ferredoxin
-
 ?
1.2.7.5 an aldehyde + H2O + oxidized ferredoxin
-
 Pyrococcus furiosus an acid + H+ + reduced ferredoxin
-
 ir
1.2.7.5 butyraldehyde + H2O + oxidized ferredoxin
-
 Pyrococcus furiosus butyrate + H+ + reduced ferredoxin
-
 ir
1.2.7.5 crotonaldehyde + H2O + oxidized ferredoxin best substrate Pyrococcus furiosus crotonate + H+ + reduced ferredoxin
-
 ir
1.2.7.5 crotonaldehyde + H2O + oxidized methyl viologen artifical electron acceptor Pyrococcus furiosus crotonate + H+ + reduced methyl viologen
-
 ir
1.2.7.5 formaldehyde + H2O + oxidized ferredoxin
-
 Pyrococcus furiosus formate + H+ + reduced ferredoxin
-
 ir
1.2.7.5 glyceraldehyde + H2O + oxidized ferredoxin
-
 Pyrococcus furiosus glycerate + H+ + reduced ferredoxin
-
 ir
1.2.7.5 glyceraldehyde + H2O + oxidized ferredoxin involved in glycolysis Pyrococcus furiosus glycerate + H+ + reduced ferredoxin
-
 ir
1.2.7.5 glyceraldehyde + H2O + oxidized methyl viologen
-
 Pyrococcus furiosus glycerate + H+ + reduced methyl viologen
-
 ir
1.2.7.5 additional information enzyme shows an active and an inactive form Pyrococcus furiosus ?
-
 ?
1.2.7.5 additional information no oxidizing activity with glyceraldehyde-3-phosphate, glyoxylate, glucose, glucose 6-phosphate, CO, H2, formate, pyruvate, 2-oxoglutarate Pyrococcus furiosus ?
-
 ?
1.2.7.5 additional information NAD(P) is no electron acceptor Pyrococcus furiosus ?
-
 ?
1.2.7.5 additional information no H2 evolution from reduced methyl viologen Pyrococcus furiosus ?
-
 ?
1.2.7.5 additional information no activity with CoA Pyrococcus furiosus ?
-
 ?
1.2.99.6 Acetaldehyde + methyl viologen
-
 Pyrococcus furiosus acetate + reduced methyl viologen
-
 ?
1.2.99.6 Butanal + methyl viologen
-
 Pyrococcus furiosus Butanoate + reduced methyl viologen
-
 ?
1.2.99.6 Crotonaldehyde + methyl viologen
-
 Pyrococcus furiosus Crotonate + reduced methyl viologen
-
 ?
1.2.99.6 Glyceraldehyde + methyl viologen
-
 Pyrococcus furiosus Glycerate + reduced methyl viologen
-
 ?
1.2.99.6 Methanal + methyl viologen
-
 Pyrococcus furiosus Formate + reduced carbamoyl methyl viologen
-
 ?
1.2.99.6 additional information its physiological role is proposed as the oxidation of glyceraldehyde to glycerate in a unique, partially nonphosphorylated glycolytic pathway that generates acetyl-CoA from glucose without participation of nicotinamide nucleotides Pyrococcus furiosus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.2.7.5 monomer 1 * 80000, SDS-PAGE Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.2.7.5 90
-
 above Pyrococcus furiosus
1.2.99.6 90
-
 above Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.2.7.5 additional information
-
 the enzyme is thermostable Pyrococcus furiosus
1.2.7.5 23
-
 pure enzyme, 5.2 mg/ml, 50 mM Tris-HCl, pH 8.0, 2 mM sodium dithionite, 2 mM DTT, 10% v/v glycerol, exposure to air, less than 20% remaining activity after 5 min, after 10 additional min 10% residual activity is reached, which stays constant for more than 30 min Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.7.5 8.4
-
 sharp maximum Pyrococcus furiosus
1.2.99.6 11
-
 activity increases up to pH 11 Pyrococcus furiosus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.2.7.5 7 8.4 activity increases with increasing pH at increasing temperature, nearly no activity below pH 7.0 Pyrococcus furiosus

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.7.5 Ferredoxin
-
 Pyrococcus furiosus
1.2.7.5 tungsten-molybdopterin tungstopterin center Pyrococcus furiosus