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Literature summary extracted from

  • Takashima, E.; Inaoka, D.K.; Osanai, A.; Nara, T.; Odaka, M.; Aoki, T.; Inaka, K.; Harada, S.; Kita, K.
    Characterization of the dihydroorotate dehydrogenase as a soluble fumarate reductase in Trypanosoma cruzi (2002), Mol. Biochem. Parasitol., 122, 189-200.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.98.1 expression in Escherichia coli strain BL21(DE3) as His-tagged enzyme Trypanosoma cruzi

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.98.1 Barbiturate dead-end inhibitor, competitive with respect to (S)-dihydroorotate Trypanosoma cruzi
1.3.98.1 Orotate competitive Trypanosoma cruzi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.98.1 0.024
-
 (S)-dihydroorotate recombinant enzyme, pH 7.5, 25°C Trypanosoma cruzi
1.3.98.1 0.026
-
 (S)-dihydroorotate native enzyme, pH 7.5, 25°C Trypanosoma cruzi
1.3.98.1 0.044
-
 fumarate recombinant enzyme, pH 7.5, 25°C Trypanosoma cruzi
1.3.98.1 0.053
-
 fumarate native enzyme, pH 7.5, 25°C Trypanosoma cruzi

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.3.5.2 cytosol
-
 Trypanosoma cruzi 5829
-
1.3.98.1 cytosol
-
 Trypanosoma cruzi 5829
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.3.5.2 34000
-
 2 * 34000, SDS-PAGE, dynamic light scattering analysis Trypanosoma cruzi
1.3.5.2 35000
-
 gel filtration Trypanosoma cruzi
1.3.98.1 34000
-
 2 * 34000, SDS-PAGE, dynamic light scattering analysis Trypanosoma cruzi
1.3.98.1 35000
-
 gel filtration Trypanosoma cruzi
1.3.98.1 36000
-
 2 * 36000, recombinant enzyme, SDS-PAGE Trypanosoma cruzi
1.3.98.1 70000
-
 about, gel filtration Trypanosoma cruzi

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.98.1 (S)-dihydroorotate + fumarate Trypanosoma cruzi enzyme is responsible for biosynthesis of succinate and orotate orotate + succinate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.5.2 Trypanosoma cruzi
-
-
-
1.3.98.1 Trypanosoma cruzi
-
-
-
1.3.98.1 Trypanosoma cruzi
-
 gene pyr4
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.5.2
-
 Trypanosoma cruzi
1.3.98.1
-
 Trypanosoma cruzi
1.3.98.1 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

EC Number Reaction Comment Organism Reaction ID
1.3.98.1 (S)-dihydroorotate + fumarate = orotate + succinate ping-pong bi-bi mechanism Trypanosoma cruzi
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.3.98.1 epimastigote
-
 Trypanosoma cruzi
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.5.2 119000
-
 cosubstrate O2, pH 7.5 Trypanosoma cruzi
1.3.5.2 245000
-
 cosubstrate 2,6-dichlorophenolindophenol, pH 7.5 Trypanosoma cruzi
1.3.5.2 548000
-
 cosubstrate Q1, pH 7.5 Trypanosoma cruzi
1.3.98.1 0.0011
-
 cytosolic fraction, native enzyme Trypanosoma cruzi
1.3.98.1 1.61
-
 purified native enzyme, electron acceptor fumarate Trypanosoma cruzi
1.3.98.1 2.95
-
 purified recombinant enzyme, electron acceptor fumarate Trypanosoma cruzi
1.3.98.1 151000
-
 cosubstrate fumarate, pH 7.5 Trypanosoma cruzi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.5.2 dihydroorotate + 2,6-dichlorophenolindophenol
-
 Trypanosoma cruzi orotate + reduced 2,6-dichlorophenolindophenol
-
 ?
1.3.5.2 additional information enzyme has methylviologen-fumarate reductase activity Trypanosoma cruzi ?
-
 ?
1.3.98.1 (S)-dihydroorotate + fumarate
-
 Trypanosoma cruzi orotate + succinate
-
 ir
1.3.98.1 (S)-dihydroorotate + fumarate enzyme is responsible for biosynthesis of succinate and orotate Trypanosoma cruzi orotate + succinate
-
 ?
1.3.98.1 (S)-dihydroorotate + O2
-
 Trypanosoma cruzi orotate + H2O2
-
 ?
1.3.98.1 dihydroorotate + 2,6-dichlorophenolindophenol
-
 Trypanosoma cruzi orotate + reduced 2,6-dichlorophenolindophenol
-
 ?
1.3.98.1 dihydroorotate + fumarate
-
 Trypanosoma cruzi orotate + ?
-
 ?
1.3.98.1 dihydroorotate + O2
-
 Trypanosoma cruzi orotate + H2O2
-
 ?
1.3.98.1 additional information enzyme has methylviologen-fumarate reductase activity Trypanosoma cruzi ?
-
 ?
1.3.98.1 orotate + ferricyanide
-
 Trypanosoma cruzi (S)-dihydroorotate + ferrocyanide
-
 ?

Subunits

EC Number Subunits Comment Organism
1.3.5.2 homodimer 2 * 34000, SDS-PAGE, dynamic light scattering analysis Trypanosoma cruzi
1.3.98.1 dimer 2 * 36000, recombinant enzyme, SDS-PAGE Trypanosoma cruzi
1.3.98.1 homodimer 2 * 34000, SDS-PAGE, dynamic light scattering analysis Trypanosoma cruzi

Synonyms

EC Number Synonyms Comment Organism
1.3.98.1 DHOD
-
 Trypanosoma cruzi
1.3.98.1 dihydroorotate dehydrogenase
-
 Trypanosoma cruzi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.98.1 25
-
 assay at Trypanosoma cruzi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.98.1 7.5
-
 assay at Trypanosoma cruzi

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.5.2 FMN
-
 Trypanosoma cruzi
1.3.98.1 FMN
-
 Trypanosoma cruzi
1.3.98.1 FMN recombinant enzyme Trypanosoma cruzi
1.3.98.1 additional information oxidized 2,6-dichlorophenol-indophenol, ferricyanide, or 1-ubiquinone can act as artificial electron acceptors Trypanosoma cruzi

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.3.98.1 0.0156
-
 Orotate versus fumarate, pH 7.5, 25°C Trypanosoma cruzi
1.3.98.1 0.0205
-
 Orotate versus (S)-dihydroorotate, pH 7.5, 25°C Trypanosoma cruzi