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Literature summary for 1.3.98.1 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)

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

Inhibitors

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 [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

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Trypanosoma cruzi 5829
-

Molecular Weight [Da]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Trypanosoma cruzi
-
-
-
Trypanosoma cruzi
-
gene pyr4
-

Purification (Commentary)

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

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

Source Tissue

Source Tissue Comment Organism Textmining
epimastigote
-
Trypanosoma cruzi
-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms

Synonyms Comment Organism
DHOD
-
Trypanosoma cruzi
dihydroorotate dehydrogenase
-
Trypanosoma cruzi

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

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 [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