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Literature summary for 1.3.98.1 extracted from
- Purification and properties of dihydroorotate oxidase from Crithidia fasciculata and Trypanosoma brucei (1983), Biochemistry, 22, 171-178.
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Orotate | - |
Crithidia fasciculata |  |
| Salicylhydroxamic acid | weak inhibition, pH 9.0, 37°C | Crithidia fasciculata | |
| suramin | weak inhibition, pH 9.0, 37°C | Crithidia fasciculata | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.001 | - |
S-dihydroorotate | pH 9.0 | Crithidia fasciculata | |
| 0.005 | - |
S-dihydroorotate | pH 7.4 | Crithidia fasciculata | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 30500 | - |
2 * 30500, SDS-PAGE | Trypanosoma brucei brucei |
| 32700 | - |
2 * 32700, SDS-PAGE | Crithidia fasciculata |
| 57000 | - |
gel filtration | Crithidia fasciculata |
| 62000 | - |
gel filtration | Trypanosoma brucei brucei |
| 63300 | - |
gel filtration | Crithidia fasciculata |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Crithidia fasciculata | - |
- |
- |
| Trypanosoma brucei brucei | - |
EATRO 110 | - |
| Trypanosoma brucei brucei EATRO 110 | - |
EATRO 110 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| acid treatment, DE-52-, Matrex Orange A-, Matrex Blue A column chromatography | Crithidia fasciculata |
| streptomycin sulfate and ammonium sulfate precipitation, DEAE-Sephacel-, affinity chromatography | Trypanosoma brucei brucei |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.63 | - |
- |
Trypanosoma brucei brucei |
| 1.4 | - |
at 37°C, 5 mM sodium diphosphate buffer | Trypanosoma brucei brucei |
| 3.99 | - |
pH 9.0, 37°C | Crithidia fasciculata |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -78°C, indefinitely stable | Crithidia fasciculata |
| 4°C, 10% loss of activity per day, full activity restored by addition of 0.1 mM 2-mercaptoethanol or dithiothreitol | Crithidia fasciculata |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-dihydroorotate + O2 | ferricyanide, 2,6-dichlorophenolindophenol, cytochrome c and quinones as alternate electron acceptor, with ferricyanide 12 times faster rate of catalysis than with air alone | Crithidia fasciculata | orotate + H2O2 | - |
? | |
| (S)-dihydroorotate + O2 | ferricyanide, 2,6-dichlorophenolindophenol, cytochrome c and quinones as alternate electron acceptor, with ferricyanide 12 times faster rate of catalysis than with air alone | Trypanosoma brucei brucei | orotate + H2O2 | - |
? | |
| (S)-dihydroorotate + O2 | ferricyanide, 2,6-dichlorophenolindophenol, cytochrome c and quinones as alternate electron acceptor, with ferricyanide 12 times faster rate of catalysis than with air alone | Trypanosoma brucei brucei EATRO 110 | orotate + H2O2 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 30500, SDS-PAGE | Trypanosoma brucei brucei |
| dimer | 2 * 32700, SDS-PAGE | Crithidia fasciculata |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
optimum for routine assay | Crithidia fasciculata |
| 37 | - |
optimum for routine assay | Trypanosoma brucei brucei |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 4 | - |
10% loss of activity per day | Crithidia fasciculata |
| 42 | - |
pH 9.0, enzyme not indefinitely stable at this temperature | Crithidia fasciculata |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 4 | - |
S-dihydroorotate | - |
Crithidia fasciculata | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.8 | - |
assay at | Crithidia fasciculata |
| 7.8 | - |
assay at | Trypanosoma brucei brucei |
| 9 | - |
standard assay at | Crithidia fasciculata |
| 9 | - |
standard assay at | Trypanosoma brucei brucei |
| 9.4 | - |
5 mM sodium diphosphate buffer | Trypanosoma brucei brucei |
| 9.5 | - |
maximum, 100 mM sodium diphosphate | Crithidia fasciculata |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 11 | best in sodium phosphate and sodium diphosphate buffer, not active below pH 6.0, 60% of activity maximum at pH 11.0 | Crithidia fasciculata |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| FMN | flavoprotein, each mol of native dimer contains 1 mol of tightly bound FMN | Crithidia fasciculata | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0045 | - |
Orotate | pH 9.0, 37°C | Crithidia fasciculata | |
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