Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli strain BL21(DE3) as His-tagged enzyme | Trypanosoma cruzi |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Barbiturate | dead-end inhibitor, competitive with respect to (S)-dihydroorotate | Trypanosoma cruzi | |
Orotate | competitive | Trypanosoma cruzi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.024 | - |
(S)-dihydroorotate | recombinant enzyme, pH 7.5, 25°C | Trypanosoma cruzi | |
0.026 | - |
(S)-dihydroorotate | native enzyme, pH 7.5, 25°C | Trypanosoma cruzi | |
0.044 | - |
fumarate | recombinant enzyme, pH 7.5, 25°C | Trypanosoma cruzi | |
0.053 | - |
fumarate | native enzyme, pH 7.5, 25°C | Trypanosoma cruzi |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Trypanosoma cruzi | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34000 | - |
2 * 34000, SDS-PAGE, dynamic light scattering analysis | Trypanosoma cruzi |
35000 | - |
gel filtration | Trypanosoma cruzi |
36000 | - |
2 * 36000, recombinant enzyme, SDS-PAGE | Trypanosoma cruzi |
70000 | - |
about, gel filtration | Trypanosoma cruzi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-dihydroorotate + fumarate | Trypanosoma cruzi | enzyme is responsible for biosynthesis of succinate and orotate | orotate + succinate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma cruzi | - |
- |
- |
Trypanosoma cruzi | - |
gene pyr4 | - |
Purification (Comment) | Organism |
---|---|
- |
Trypanosoma cruzi |
native enzyme from the cytosolic fraction by gel filtration and immunoaffinity chromatography, recombinant His-tagged enzyme by nickel affinity chromatography to homogeneity and subsequent clevage of the tag by thrombin | Trypanosoma cruzi |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(S)-dihydroorotate + fumarate = orotate + succinate | ping-pong bi-bi mechanism | Trypanosoma cruzi |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
epimastigote | - |
Trypanosoma cruzi | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0011 | - |
cytosolic fraction, native enzyme | Trypanosoma cruzi |
1.61 | - |
purified native enzyme, electron acceptor fumarate | Trypanosoma cruzi |
2.95 | - |
purified recombinant enzyme, electron acceptor fumarate | Trypanosoma cruzi |
151000 | - |
cosubstrate fumarate, pH 7.5 | Trypanosoma cruzi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-dihydroorotate + fumarate | - |
Trypanosoma cruzi | orotate + succinate | - |
ir | |
(S)-dihydroorotate + fumarate | enzyme is responsible for biosynthesis of succinate and orotate | Trypanosoma cruzi | orotate + succinate | - |
? | |
(S)-dihydroorotate + O2 | - |
Trypanosoma cruzi | orotate + H2O2 | - |
? | |
dihydroorotate + 2,6-dichlorophenolindophenol | - |
Trypanosoma cruzi | orotate + reduced 2,6-dichlorophenolindophenol | - |
? | |
dihydroorotate + fumarate | - |
Trypanosoma cruzi | orotate + ? | - |
? | |
dihydroorotate + O2 | - |
Trypanosoma cruzi | orotate + H2O2 | - |
? | |
additional information | enzyme has methylviologen-fumarate reductase activity | Trypanosoma cruzi | ? | - |
? | |
orotate + ferricyanide | - |
Trypanosoma cruzi | (S)-dihydroorotate + ferrocyanide | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 36000, recombinant enzyme, SDS-PAGE | Trypanosoma cruzi |
homodimer | 2 * 34000, SDS-PAGE, dynamic light scattering analysis | Trypanosoma cruzi |
Synonyms | Comment | Organism |
---|---|---|
DHOD | - |
Trypanosoma cruzi |
dihydroorotate dehydrogenase | - |
Trypanosoma cruzi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Trypanosoma cruzi |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Trypanosoma cruzi |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | - |
Trypanosoma cruzi | |
FMN | recombinant enzyme | Trypanosoma cruzi | |
additional information | oxidized 2,6-dichlorophenol-indophenol, ferricyanide, or 1-ubiquinone can act as artificial electron acceptors | Trypanosoma cruzi |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0156 | - |
Orotate | versus fumarate, pH 7.5, 25°C | Trypanosoma cruzi | |
0.0205 | - |
Orotate | versus (S)-dihydroorotate, pH 7.5, 25°C | Trypanosoma cruzi |