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Literature summary for 1.2.7.11 extracted from

  • Fukuda, E.; Kino, H.; Matsuzawa, H.; Wakagi, T.
    Role of a highly conserved YPITP motif in 2-oxoacid:ferredoxin oxidoreductase: heterologous expression of the gene from Sulfolobus sp.strain 7, and characterization of the recombinant and variant enzymes (2001), Eur. J. Biochem., 268, 5639-5646.
    View publication on PubMed
Search for more literature for 1.2.7.11

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli, wild-type recombinant enzyme is indistinguishable from the natural one in every criterion investigated Sulfolobus sp.

Protein Variants

Protein Variants Comment Organism
I255L kcat/Km for pyruvate is 11% of wild-type value, kcat/KM for 2-oxoglutarate is 21% of wild-type value Sulfolobus sp.
I255M kcat/Km for pyruvate is 13% of wild-type value, kcat/KM for 2-oxoglutarate is 2% of wild-type value Sulfolobus sp.
I255S kcat/Km for pyruvate is 23% of wild-type value, kcat/KM for 2-oxoglutarate is 35% of wild-type value Sulfolobus sp.
I255V kcat/Km for pyruvate is 14% of wild-type value, kcat/KM for 2-oxoglutarate is 38% of wild-type value Sulfolobus sp.
P254G kcat/Km for pyruvate is 12% of wild-type value, kcat/KM for 2-oxoglutarate is 20% of wild-type value Sulfolobus sp.
P257A no enzyme activity at either 50 or 80°C Sulfolobus sp.
P257G no enzyme activity at either 50 or 80°C Sulfolobus sp.
P257V no enzyme activity at either 50 or 80°C Sulfolobus sp.
T256A kcat and Km for 2-oxoglutarate are 33% and 51%, respectively, as compared with that of the wild-type enzyme Sulfolobus sp.
T256A kcat/Km for pyruvate is 21% of wild-type value, kcat/KM for 2-oxoglutarate is 15% of wild-type value Sulfolobus sp.
T256S kcat/Km for pyruvate is 15% of wild-type value, kcat/KM for 2-oxoglutarate is 92% of wild-type value Sulfolobus sp.
T256V kcat/Km for pyruvate is 17% of wild-type value, kcat/KM for 2-oxoglutarate is 68% of wild-type value Sulfolobus sp.
Y253A no enzyme activity at either 50 or 80°C Sulfolobus sp.
Y253F kcat/Km for pyruvate is 3.9% of wild-type value, kcat/KM for 2-oxoglutarate is 16% of wild-type value Sulfolobus sp.
Y253W no enzyme activity at either 50 or 80°C Sulfolobus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.11
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256S Sulfolobus sp.
0.13
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255S Sulfolobus sp.
0.14
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256A Sulfolobus sp.
0.17
-
 pyruvate pH 7.5, 80°C, mutant enzyme P254G Sulfolobus sp.
0.17
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256V Sulfolobus sp.
0.19
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255M Sulfolobus sp.
0.25
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255V Sulfolobus sp.
0.28
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255L Sulfolobus sp.
0.28
-
 pyruvate pH 7.5, 80°C, naturale enzyme Sulfolobus sp.
0.31
-
 pyruvate pH 7.5, 80°C, mutant enzyme P253F Sulfolobus sp.
0.33
-
 pyruvate pH 7.5, 80°C, recombinant enzyme Sulfolobus sp.
0.33
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256V Sulfolobus sp.
0.37
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256A Sulfolobus sp.
0.72
-
 2-oxoglutarate pH 7.5, 80°C, recombinant enzyme Sulfolobus sp.
0.85
-
 2-oxoglutarate pH 7.5, 80°C, natural enzyme Sulfolobus sp.
0.89
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255S Sulfolobus sp.
1.1
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255L Sulfolobus sp.
1.2
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme P254G Sulfolobus sp.
1.3
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255M Sulfolobus sp.
1.5
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme P253F Sulfolobus sp.
2.1
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256S Sulfolobus sp.
3.3
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255V Sulfolobus sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ natural enzyme: 1.4 mol/mol, recombinant enzyme 1.9 mol/mol Sulfolobus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34000
-
 1 * 71000 (subunit a) + 1 * 34000 (subunit b), SDS-PAGE Sulfolobus sp.
71000
-
 1 * 71000 (subunit a) + 1 * 34000 (subunit b), SDS-PAGE Sulfolobus sp.
105000
-
 gel filtration Sulfolobus sp.

Organism

Organism UniProt Comment Textmining
Sulfolobus sp.
-
-
-
Sulfolobus sp. 7
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Sulfolobus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxoglutarate + CoA + 2 oxidized ferredoxin kcat/Km for 2-oxobutyrate is 12% of the kcat/Km-value for pyruvate, a cognate Zn-7Fe-ferredoxin serves as an electron acceptor Sulfolobus sp. succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
 ?
2-oxoglutarate + CoA + 2 oxidized ferredoxin kcat/Km for 2-oxobutyrate is 12% of the kcat/Km-value for pyruvate, a cognate Zn-7Fe-ferredoxin serves as an electron acceptor Sulfolobus sp. 7 succinyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
 ?
additional information the YPITP-motif is essential for the turnover of the reaction rather than the affinity toward 2-oxoacid Sulfolobus sp. ?
-
 ?
additional information the YPITP-motif is essential for the turnover of the reaction rather than the affinity toward 2-oxoacid Sulfolobus sp. 7 ?
-
 ?
pyruvate + CoA + 2 oxidized ferredoxin a cognate Zn-7Fe-ferredoxin serves as an electron acceptor Sulfolobus sp. acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
 ?
pyruvate + CoA + 2 oxidized ferredoxin a cognate Zn-7Fe-ferredoxin serves as an electron acceptor Sulfolobus sp. 7 acetyl-CoA + CO2 + 2 reduced ferredoxin + 2 H+
-
 ?

Subunits

Subunits Comment Organism
dimer 1 * 71000 (subunit a) + 1 * 34000 (subunit b), SDS-PAGE Sulfolobus sp.

Synonyms

Synonyms Comment Organism
OFOR
-
 Sulfolobus sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
 assay at Sulfolobus sp.
90
-
-
 Sulfolobus sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
 pH 7.2, 30 min, about 20% loss of activity, natural and recombinant wild-type enzyme Sulfolobus sp.
80
-
 pH 7.2, 30 min, about 40% loss of activity, natural and recombinant wild-type enzyme Sulfolobus sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.7
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255V Sulfolobus sp.
2.2
-
 pyruvate pH 7.5, 80°C, mutant enzyme P253F Sulfolobus sp.
3.2
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255S Sulfolobus sp.
3.6
-
 pyruvate pH 7.5, 80°C, mutant enzyme P254G Sulfolobus sp.
4.1
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256S Sulfolobus sp.
4.3
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256A Sulfolobus sp.
4.6
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256V Sulfolobus sp.
5.1
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255L Sulfolobus sp.
5.2
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme P254G Sulfolobus sp.
5.3
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme P253F Sulfolobus sp.
5.5
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256A Sulfolobus sp.
5.7
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255L Sulfolobus sp.
6
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255V Sulfolobus sp.
6.7
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256V Sulfolobus sp.
7.4
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255S Sulfolobus sp.
7.4
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256S Sulfolobus sp.
7.9
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255M Sulfolobus sp.
10
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255M Sulfolobus sp.
17
-
 2-oxoglutarate pH 7.5, 80°C, recombinant enzyme Sulfolobus sp.
19
-
 2-oxoglutarate pH 7.5, 80°C, natural enzyme Sulfolobus sp.
48
-
 pyruvate pH 7.5, 80°C, recombinant enzyme Sulfolobus sp.
51
-
 pyruvate pH 7.5, 80°C, naturale enzyme Sulfolobus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Sulfolobus sp.
8.5
-
-
 Sulfolobus sp.

Cofactor

Cofactor Comment Organism Structure
thiamine diphosphate
-
 Sulfolobus sp.

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.52
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255V Sulfolobus sp.
3.4
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256S Sulfolobus sp.
3.6
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme P253F Sulfolobus sp.
4.3
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme P254G Sulfolobus sp.
4.6
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255L Sulfolobus sp.
7.2
-
 pyruvate pH 7.5, 80°C, mutant enzyme P253F Sulfolobus sp.
7.7
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255M Sulfolobus sp.
8.3
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme I255S Sulfolobus sp.
15
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256A Sulfolobus sp.
20
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255L Sulfolobus sp.
20
-
 2-oxoglutarate pH 7.5, 80°C, mutant enzyme T256V Sulfolobus sp.
21
-
 pyruvate pH 7.5, 80°C, mutant enzyme P254G Sulfolobus sp.
22
-
 2-oxoglutarate pH 7.5, 80°C, natural enzyme Sulfolobus sp.
23
-
 2-oxoglutarate pH 7.5, 80°C, recombinant enzyme Sulfolobus sp.
24
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255V Sulfolobus sp.
25
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255S Sulfolobus sp.
28
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256V Sulfolobus sp.
31
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256A Sulfolobus sp.
38
-
 pyruvate pH 7.5, 80°C, mutant enzyme T256S Sulfolobus sp.
41
-
 pyruvate pH 7.5, 80°C, mutant enzyme I255M Sulfolobus sp.
146
-
 pyruvate pH 7.5, 80°C, recombinant enzyme Sulfolobus sp.
182
-
 pyruvate pH 7.5, 80°C, naturale enzyme Sulfolobus sp.