BRENDA - Enzyme Database show

Crystal structures of archaeal 2-oxoacid:ferredoxin oxidoreductases from Sulfolobus tokodaii

Yan, Z.; Maruyama, A.; Arakawa, T.; Fushinobu, S.; Wakagi, T.; Sci. Rep. 6, 33061 (2016)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.2.7.11
expressed in Escherichia coli C43 (DE3) cells; expressed in Escherichia coli C43 (DE3) cells; expression in Escherichia coli (DE3); expression in Escherichia coli (DE3)
Sulfurisphaera tokodaii
Crystallization (Commentary)
EC Number
Crystallization
Organism
1.2.7.11
crystals are grown at 20C using sitting drop vapor diffusion. The structure of the recombinant enzyme StOFOR2 by the single-wavelength anomalous dispersion method is solved using a selenomethionine(SeMet)-labeled protein crystal, and the structures of the ligand-free (2.1 resolution) and pyruvate-complexed (2.2 ) forms are determined. In the structure of the recombinant enzyme StOFOR2 in unreacted pyruvate complex form, the carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus. A possible complex structural model is constructed by placing a Zn2+-containing dicluster ferredoxin of Sulfolobus tokodaii into the large pocket of the recombinant StOFOR2 enzyme, providing insight into the electron transfer between the two redox proteins; crystals of the StOFOR1 enzyme are prepared by co-crystallization with 50 mM 2-oxobutyrate and 1 mM CoA. Crystals are grown at 25C using sitting drop vapor diffusion. In the structure of StOFOR1 co-crystallized with 2-oxobutyrate, electron density corresponding to a 1-hydroxypropyl group (post-decarboxylation state) is observed at the thiazole ring of thiamine diphosphate. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus; sitting drop vapor diffusion method, using 0.7 M ammonium tartrate dibasic and 0.1 M Tris-HCl (pH 8.5); sitting drop vapor diffusion method, using 15% (w/v) PEG3350, 0.1 M Tris-HCl (pH 8.5) and 0.1 M NaBr
Sulfurisphaera tokodaii
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.2.7.11
D468A
inactive with 2-oxoglutarate and pyruvate as substrate; mutant enzyme StOFOR1 with mutation D468A in alpha-subunit. Vmax with pyruvate as substrate is 1.3% compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
1.2.7.11
K49I
inactive with 2-oxoglutarate as substrate; mutant enzyme StOFOR1 with mutation K49I in alpha-subunit. Vmax with pyruvate as substrate is 28% compared to wild-type enzyme, Km with pyruvate as substrate is 2.8fold higher as compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
1.2.7.11
S41A
mutant enzyme StOFOR1 with mutation S41A in alpha-subunit. Vmax with pyruvate as substrate is 29% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 40% compared to wild-type enzyme, Km with pyruvate as substrate is 1.5fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.5fold higher as compared to wild-type enzyme; the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
1.2.7.11
T349L
mutant enzyme StOFOR1 with mutation T349L in alpha-subunit. Vmax with pyruvate as substrate is 43% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 74% compared to wild-type enzyme, Km with pyruvate as substrate is 1.6fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.1fold higher as compared to wild-type enzyme; the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.2.7.11
0.32
-
pyruvate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1; wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
0.49
-
pyruvate
mutant enzyme S41A, at pH 8.5 and 80C; pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
0.51
-
pyruvate
mutant enzyme T349L, at pH 8.5 and 80C; pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
0.91
-
pyruvate
mutant enzyme K49I, at pH 8.5 and 80C; pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation K49I in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
1.6
-
pyruvate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2; wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
2.1
-
2-oxoglutarate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1; wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
2.3
-
2-oxoglutarate
mutant enzyme T349L, at pH 8.5 and 80C; pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
3.2
-
2-oxoglutarate
mutant enzyme S41A, at pH 8.5 and 80C; pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
15
-
2-oxoglutarate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2; wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.2.7.11
Mg2+
required; required
Sulfurisphaera tokodaii
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.2.7.11
34000
-
;
Sulfurisphaera tokodaii
1.2.7.11
70000
-
;
Sulfurisphaera tokodaii
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii 7
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.2.7.11
Sulfurisphaera tokodaii
Q96XT2 and Q96XT4
Q96XT2: alpha-subunit (ST2435), Q96XT4: beta-subunit (ST2433); there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. Recombinant protein samples of STK_24350/STK_24330 are prepared and this enzyme is designated as StOFOR2
-
1.2.7.11
Sulfurisphaera tokodaii
Q96XT4
beta-subunit
-
1.2.7.11
Sulfurisphaera tokodaii
Q96Y66 and Q96Y68
Q96Y66: alpha-subunit (ST2300), Q96Y68: beta-subunit (ST2298)
-
1.2.7.11
Sulfurisphaera tokodaii
Q96Y68
beta-subunit
-
1.2.7.11
Sulfurisphaera tokodaii 7
Q96XT4
beta-subunit
-
1.2.7.11
Sulfurisphaera tokodaii 7
Q96Y68
beta-subunit
-
1.2.7.11
Sulfurisphaera tokodaii DSM 16993
Q96XT2 and Q96XT4
Q96XT2: alpha-subunit (ST2435), Q96XT4: beta-subunit (ST2433); there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. Recombinant protein samples of STK_24350/STK_24330 are prepared and this enzyme is designated as StOFOR2
-
1.2.7.11
Sulfurisphaera tokodaii DSM 16993
Q96Y66 and Q96Y68
Q96Y66: alpha-subunit (ST2300), Q96Y68: beta-subunit (ST2298)
-
Purification (Commentary)
EC Number
Commentary
Organism
1.2.7.11
DEAE Sepharose column chromatography and Superdex 200 gel filtration; DEAE Sepharose column chromatography and Superdex 200 gel filtration
Sulfurisphaera tokodaii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.7.11
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
1.2.7.11
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
1.2.7.11
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
?
-
-
-
?
1.2.7.11
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.2.7.11
heterodimer
1 * 70000 + 1 * 34000, SDS-PAGE; 1 * 70000 + 1 * 34000, SDS-PAGE; crystals consist of two protomers (1 and 2), each of which consists of one heterodimer of alpha- and beta-subunits; crystals consist of two protomers (1 and 2), each of which consists of one heterodimer of alpha- and beta-subunits
Sulfurisphaera tokodaii
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.2.7.11
80
-
assay at; assay at
Sulfurisphaera tokodaii
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.2.7.11
8.5
-
assay at; assay at
Sulfurisphaera tokodaii
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.2.7.11
Ferredoxin
alpha/beta-subunit heterodimers lack an intramolecular ferredoxin domain. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule; alpha/beta-subunit heterodimers lack an intramolecular ferredoxin domain. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
Sulfurisphaera tokodaii
1.2.7.11
thiamine diphosphate
alpha/beta-subunit heterodimers contain thiamin diphosphate; alpha/beta-subunit heterodimers contain thiamin diphosphate
Sulfurisphaera tokodaii
1.2.7.11
[4Fe-4S]-center
alpha/beta-subunit heterodimers contain 4Fe-4S cluster; alpha/beta-subunit heterodimers contain 4Fe-4S cluster
Sulfurisphaera tokodaii
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.2.7.11
expressed in Escherichia coli C43 (DE3) cells
Sulfurisphaera tokodaii
1.2.7.11
expression in Escherichia coli (DE3)
Sulfurisphaera tokodaii
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.2.7.11
Ferredoxin
alpha/beta-subunit heterodimers lack an intramolecular ferredoxin domain. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
Sulfurisphaera tokodaii
1.2.7.11
thiamine diphosphate
alpha/beta-subunit heterodimers contain thiamin diphosphate
Sulfurisphaera tokodaii
1.2.7.11
thiamine diphosphate
-
Sulfurisphaera tokodaii
1.2.7.11
[4Fe-4S]-center
alpha/beta-subunit heterodimers contain 4Fe-4S cluster
Sulfurisphaera tokodaii
1.2.7.11
[4Fe-4S]-center
-
Sulfurisphaera tokodaii
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.2.7.11
crystals are grown at 20C using sitting drop vapor diffusion. The structure of the recombinant enzyme StOFOR2 by the single-wavelength anomalous dispersion method is solved using a selenomethionine(SeMet)-labeled protein crystal, and the structures of the ligand-free (2.1 resolution) and pyruvate-complexed (2.2 ) forms are determined. In the structure of the recombinant enzyme StOFOR2 in unreacted pyruvate complex form, the carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus. A possible complex structural model is constructed by placing a Zn2+-containing dicluster ferredoxin of Sulfolobus tokodaii into the large pocket of the recombinant StOFOR2 enzyme, providing insight into the electron transfer between the two redox proteins
Sulfurisphaera tokodaii
1.2.7.11
crystals of the StOFOR1 enzyme are prepared by co-crystallization with 50 mM 2-oxobutyrate and 1 mM CoA. Crystals are grown at 25C using sitting drop vapor diffusion. In the structure of StOFOR1 co-crystallized with 2-oxobutyrate, electron density corresponding to a 1-hydroxypropyl group (post-decarboxylation state) is observed at the thiazole ring of thiamine diphosphate. The binding pockets of the 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii surrounding the methyl or propyl group of the ligands are wider than that of 2-oxoacid oxidoreductase enzymes from Desulfovbrio africanus
Sulfurisphaera tokodaii
1.2.7.11
sitting drop vapor diffusion method, using 0.7 M ammonium tartrate dibasic and 0.1 M Tris-HCl (pH 8.5)
Sulfurisphaera tokodaii
1.2.7.11
sitting drop vapor diffusion method, using 15% (w/v) PEG3350, 0.1 M Tris-HCl (pH 8.5) and 0.1 M NaBr
Sulfurisphaera tokodaii
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.2.7.11
D468A
mutant enzyme StOFOR1 with mutation D468A in alpha-subunit. Vmax with pyruvate as substrate is 1.3% compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
1.2.7.11
D468A
inactive with 2-oxoglutarate and pyruvate as substrate
Sulfurisphaera tokodaii
1.2.7.11
K49I
mutant enzyme StOFOR1 with mutation K49I in alpha-subunit. Vmax with pyruvate as substrate is 28% compared to wild-type enzyme, Km with pyruvate as substrate is 2.8fold higher as compared to wild-type enzyme. No activity is detected with 2-oxoglutarate
Sulfurisphaera tokodaii
1.2.7.11
K49I
inactive with 2-oxoglutarate as substrate
Sulfurisphaera tokodaii
1.2.7.11
S41A
mutant enzyme StOFOR1 with mutation S41A in alpha-subunit. Vmax with pyruvate as substrate is 29% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 40% compared to wild-type enzyme, Km with pyruvate as substrate is 1.5fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.5fold higher as compared to wild-type enzyme
Sulfurisphaera tokodaii
1.2.7.11
S41A
the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
1.2.7.11
T349L
mutant enzyme StOFOR1 with mutation T349L in alpha-subunit. Vmax with pyruvate as substrate is 43% compared to wild-type enzyme, Vmax with 2-oxoglutarate as substrate is 74% compared to wild-type enzyme, Km with pyruvate as substrate is 1.6fold higher as compared to wild-type enzyme, Km with 2-oxoglutarate as substrate is 1.1fold higher as compared to wild-type enzyme
Sulfurisphaera tokodaii
1.2.7.11
T349L
the mutant shows reduced activity compared to the wild type enzyme
Sulfurisphaera tokodaii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.2.7.11
0.32
-
pyruvate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1
Sulfurisphaera tokodaii
1.2.7.11
0.32
-
pyruvate
wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
0.49
-
pyruvate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
0.49
-
pyruvate
mutant enzyme S41A, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
0.51
-
pyruvate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
0.51
-
pyruvate
mutant enzyme T349L, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
0.91
-
pyruvate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation K49I in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
0.91
-
pyruvate
mutant enzyme K49I, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
1.6
-
pyruvate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2
Sulfurisphaera tokodaii
1.2.7.11
1.6
-
pyruvate
wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
2.1
-
2-oxoglutarate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR1
Sulfurisphaera tokodaii
1.2.7.11
2.1
-
2-oxoglutarate
wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
2.3
-
2-oxoglutarate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation T349L in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
2.3
-
2-oxoglutarate
mutant enzyme T349L, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
3.2
-
2-oxoglutarate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, mutant enzyme StOFOR1 with mutation S41A in alpha-subunit
Sulfurisphaera tokodaii
1.2.7.11
3.2
-
2-oxoglutarate
mutant enzyme S41A, at pH 8.5 and 80C
Sulfurisphaera tokodaii
1.2.7.11
15
-
2-oxoglutarate
pH 8.5, 80C, cosubstrate: oxidized methyl viologen, enzyme StOFOR2
Sulfurisphaera tokodaii
1.2.7.11
15
-
2-oxoglutarate
wild type enzyme, at pH 8.5 and 80C
Sulfurisphaera tokodaii
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.2.7.11
Mg2+
required
Sulfurisphaera tokodaii
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.2.7.11
34000
-
-
Sulfurisphaera tokodaii
1.2.7.11
70000
-
-
Sulfurisphaera tokodaii
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
Sulfurisphaera tokodaii 7
-
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.2.7.11
-
Sulfurisphaera tokodaii
1.2.7.11
DEAE Sepharose column chromatography and Superdex 200 gel filtration
Sulfurisphaera tokodaii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.2.7.11
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
1.2.7.11
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
1.2.7.11
2-oxobutyrate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
propanoyl-CoA + CO2 + 2 reduced methyl viologen
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. Because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized ferredoxin
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced ferredoxin + H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
1.2.7.11
2-oxoglutarate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
succinyl-CoA + CO2 + reduced methyl viologen + H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
the recombinant enzyme StOFOR2 exhibits a preference for pyruvate over 2-oxoglutarate. The carboxylate group of pyruvate is recognized by Arg344 and Thr257 from the alpha-subunit of
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized ferredoxin
because 2-oxoacid oxidoreductase enzymes from Sulfolobus tokodaii lack the intramolecular Fd-like domain V, there is a large pocket surrounded by domains III and VI in each protomer, which appears to be able to bind an external ferredoxin molecule
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + 2 oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii DSM 16993
acetyl-CoA + CO2 + 2 reduced methyl viologen + 2 H+
-
-
-
?
1.2.7.11
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii
?
-
-
-
?
1.2.7.11
pyruvate + CoA + oxidized methyl viologen
-
739701
Sulfurisphaera tokodaii 7
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.2.7.11
heterodimer
crystals consist of two protomers (1 and 2), each of which consists of one heterodimer of alpha- and beta-subunits
Sulfurisphaera tokodaii
1.2.7.11
heterodimer
1 * 70000 + 1 * 34000, SDS-PAGE
Sulfurisphaera tokodaii
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.2.7.11
80
-
assay at
Sulfurisphaera tokodaii
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.2.7.11
8.5
-
assay at
Sulfurisphaera tokodaii
General Information
EC Number
General Information
Commentary
Organism
1.2.7.11
metabolism
can utilize both pyruvate and 2-oxoglutarate, playing a key role in the central metabolism; there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. It may be expressed in vivo under some culture conditions
Sulfurisphaera tokodaii
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.2.7.11
metabolism
can utilize both pyruvate and 2-oxoglutarate, playing a key role in the central metabolism
Sulfurisphaera tokodaii
1.2.7.11
metabolism
there is no evidence for the expression of the StOFOR2 (tk_24350/stk_24330 genes) in Sulfolobus tokodaii. It may be expressed in vivo under some culture conditions
Sulfurisphaera tokodaii