BRENDA - Enzyme Database show

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

Mukund, S.; Adams, M.W.W.; J. Biol. Chem. 266, 14208-14216 (1991)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
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
Commentary
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
Commentary
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
Commentary
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
1.0 gatoms per 85000g of protein; part of a tungsten-molybdopterin cofactor
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]
Commentary
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
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.2.7.5
an aldehyde + H2O + oxidized ferredoxin
Pyrococcus furiosus
-
an acid + H+ + reduced ferredoxin
-
Pyrococcus furiosus
?
1.2.7.5
glyceraldehyde + H2O + oxidized ferredoxin
Pyrococcus furiosus
involved in glycolysis
glycerate + H+ + reduced ferredoxin
-
Pyrococcus furiosus
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
Primary Accession No. (UniProt)
Commentary
Textmining
1.2.7.5
Pyrococcus furiosus
-
hyperthermophilic archaeon; 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
Commentary
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]
Commentary
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
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.7.5
acetaldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
acetate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
an aldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
an acid + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
?
1.2.7.5
an aldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
an acid + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
butyraldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
butyrate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
crotonaldehyde + H2O + oxidized ferredoxin
best substrate
11938
Pyrococcus furiosus
pyruvate + H+ + reduced ferredoxin
-
-
-
ir
1.2.7.5
crotonaldehyde + H2O + oxidized methyl viologen
artifical electron acceptor
11938
Pyrococcus furiosus
pyruvate + H+ + reduced methyl viologen
-
-
-
ir
1.2.7.5
formaldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
formate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
glyceraldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
glycerate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
glyceraldehyde + H2O + oxidized ferredoxin
involved in glycolysis
11938
Pyrococcus furiosus
glycerate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
glyceraldehyde + H2O + oxidized methyl viologen
-
11938
Pyrococcus furiosus
glycerate + H+ + reduced methyl viologen
-
11938
Pyrococcus furiosus
ir
1.2.7.5
additional information
enzyme shows an active and an inactive form
11938
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
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.7.5
additional information
NAD(P) is no electron acceptor
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.7.5
additional information
no H2 evolution from reduced methyl viologen
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.7.5
additional information
no activity with CoA
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.99.6
Acetaldehyde + methyl viologen
-
11938
Pyrococcus furiosus
acetate + reduced methyl viologen
-
-
-
-
1.2.99.6
Butanal + methyl viologen
-
11938
Pyrococcus furiosus
Butanoate + reduced methyl viologen
-
-
-
-
1.2.99.6
Crotonaldehyde + methyl viologen
-
11938
Pyrococcus furiosus
Crotonate + reduced methyl viologen
-
-
-
-
1.2.99.6
Glyceraldehyde + methyl viologen
-
11938
Pyrococcus furiosus
Glycerate + reduced methyl viologen
-
-
-
-
1.2.99.6
Methanal + methyl viologen
-
11938
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
11938
Pyrococcus furiosus
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.2.7.5
monomer
1 * 80000, SDS-PAGE
Pyrococcus furiosus
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
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]
Commentary
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
Commentary
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
Commentary
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
Commentary
Organism
Structure
1.2.7.5
Ferredoxin
-
Pyrococcus furiosus
1.2.7.5
tungsten-molybdopterin
tungstopterin center
Pyrococcus furiosus
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.2.7.5
additional information
no effect of CoA
Pyrococcus furiosus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.2.7.5
Ferredoxin
-
Pyrococcus furiosus
1.2.7.5
tungsten-molybdopterin
tungstopterin center
Pyrococcus furiosus
General Stability (protein specific)
EC Number
General Stability
Organism
1.2.7.5
10% glycerol and 2 mM DTT stabilize during purification
Pyrococcus furiosus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
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] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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 (protein specific)
EC Number
Metals/Ions
Commentary
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
1.0 gatoms per 85000g of protein; part of a tungsten-molybdopterin cofactor
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] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
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) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.2.7.5
an aldehyde + H2O + oxidized ferredoxin
Pyrococcus furiosus
-
an acid + H+ + reduced ferredoxin
-
Pyrococcus furiosus
?
1.2.7.5
glyceraldehyde + H2O + oxidized ferredoxin
Pyrococcus furiosus
involved in glycolysis
glycerate + H+ + reduced ferredoxin
-
Pyrococcus furiosus
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
?
-
-
-
Oxidation Stability (protein specific)
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) (protein specific)
EC Number
Commentary
Organism
1.2.7.5
to homogeneity under strict anaerobic conditions
Pyrococcus furiosus
1.2.99.6
-
Pyrococcus furiosus
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
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 (protein specific)
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) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.7.5
acetaldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
acetate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
an aldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
an acid + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
?
1.2.7.5
an aldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
an acid + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
butyraldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
butyrate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
crotonaldehyde + H2O + oxidized ferredoxin
best substrate
11938
Pyrococcus furiosus
pyruvate + H+ + reduced ferredoxin
-
-
-
ir
1.2.7.5
crotonaldehyde + H2O + oxidized methyl viologen
artifical electron acceptor
11938
Pyrococcus furiosus
pyruvate + H+ + reduced methyl viologen
-
-
-
ir
1.2.7.5
formaldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
formate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
glyceraldehyde + H2O + oxidized ferredoxin
-
11938
Pyrococcus furiosus
glycerate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
glyceraldehyde + H2O + oxidized ferredoxin
involved in glycolysis
11938
Pyrococcus furiosus
glycerate + H+ + reduced ferredoxin
-
11938
Pyrococcus furiosus
ir
1.2.7.5
glyceraldehyde + H2O + oxidized methyl viologen
-
11938
Pyrococcus furiosus
glycerate + H+ + reduced methyl viologen
-
11938
Pyrococcus furiosus
ir
1.2.7.5
additional information
enzyme shows an active and an inactive form
11938
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
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.7.5
additional information
NAD(P) is no electron acceptor
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.7.5
additional information
no H2 evolution from reduced methyl viologen
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.7.5
additional information
no activity with CoA
11938
Pyrococcus furiosus
?
-
-
-
-
1.2.99.6
Acetaldehyde + methyl viologen
-
11938
Pyrococcus furiosus
acetate + reduced methyl viologen
-
-
-
-
1.2.99.6
Butanal + methyl viologen
-
11938
Pyrococcus furiosus
Butanoate + reduced methyl viologen
-
-
-
-
1.2.99.6
Crotonaldehyde + methyl viologen
-
11938
Pyrococcus furiosus
Crotonate + reduced methyl viologen
-
-
-
-
1.2.99.6
Glyceraldehyde + methyl viologen
-
11938
Pyrococcus furiosus
Glycerate + reduced methyl viologen
-
-
-
-
1.2.99.6
Methanal + methyl viologen
-
11938
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
11938
Pyrococcus furiosus
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.2.7.5
monomer
1 * 80000, SDS-PAGE
Pyrococcus furiosus
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.2.7.5
90
-
above
Pyrococcus furiosus
1.2.99.6
90
-
above
Pyrococcus furiosus
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
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 (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
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 (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
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