Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | activates | Faecalicatena orotica | |
cysteine | L-cysteine activates strongly, D-cysteine is also an activator, 1.8 mM L-cysteine is required at pH 8 for half-maximal activity, 10 mM for near-maximal activity | Faecalicatena orotica | |
dithiothreitol | activates | Faecalicatena orotica | |
glutathione | activates | Faecalicatena orotica | |
Mercaptoacetic acid | activates | Faecalicatena orotica | |
additional information | thiol activation involves reduction of side chain of a cysteine residue corresponding to Cys-130, which functions as a general base | Faecalicatena orotica | |
additional information | not activated by L-serine or L-alanine | Faecalicatena orotica |
Cloned (Comment) | Organism |
---|---|
genes encoding the two alpha and beta polypeptides, expression in Escherichia coli | Bacillus subtilis |
genes encoding the two alpha and beta polypeptides, expression in Escherichia coli | Lactococcus lactis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.09 | - |
L-dihydroorotate | pH 8, 25°C | Faecalicatena orotica | |
0.13 | - |
NAD+ | pH 8, 25°C | Faecalicatena orotica |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Bacillus subtilis | 5829 | - |
cytosol | - |
Lactococcus lactis | 5829 | - |
cytosol | - |
Enterococcus faecalis | 5829 | - |
cytosol | - |
Faecalicatena orotica | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28000 | - |
alpha2,beta2, 2 * 32000 + 2 * 28000, SDS-PAGE | Faecalicatena orotica |
32000 | - |
alpha2,beta2, 2 * 32000 + 2 * 28000, SDS-PAGE | Faecalicatena orotica |
120000 | - |
analytical gel filtration | Faecalicatena orotica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dihydroorotate + NAD+ | Lactococcus lactis | fourth step and sole redox reaction in the pyrimidine de novo biosynthetic pathway | orotate + NADH + H+ | - |
r | |
dihydroorotate + NAD+ | Enterococcus faecalis | fourth step and sole redox reaction in the pyrimidine de novo biosynthetic pathway | orotate + NADH + H+ | - |
r | |
dihydroorotate + NAD+ | Faecalicatena orotica | fourth step and sole redox reaction in the pyrimidine de novo biosynthetic pathway | orotate + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
- |
- |
Enterococcus faecalis | - |
- |
- |
Faecalicatena orotica | - |
identical with Clostridium oroticum | - |
Lactococcus lactis | - |
family IB enzyme, also contains family IA enzyme | - |
Purification (Comment) | Organism |
---|---|
- |
Faecalicatena orotica |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(S)-dihydroorotate + NAD+ = orotate + NADH + H+ | electron transfer mechanism | Lactococcus lactis | |
(S)-dihydroorotate + NAD+ = orotate + NADH + H+ | ping-pong steady-state kinetic mechanism | Enterococcus faecalis | |
(S)-dihydroorotate + NAD+ = orotate + NADH + H+ | ping-pong steady-state kinetic mechanism | Faecalicatena orotica |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Faecalicatena orotica |
Storage Stability | Organism |
---|---|
-20°C, 0.27 M sodium phosphate, pH 6.3, over 6 months, less than 15% loss of activity | Faecalicatena orotica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-dihydroorotate + NAD+ | equilibrium favours direction of orotate reduction | Faecalicatena orotica | orotate + NADH + H+ | - |
? | |
(S)-dihydroorotate + NAD+ | equilibrium favours direction of orotate reduction | Faecalicatena orotica | orotate + NADH + H+ | - |
r | |
(S)-dihydroorotate + NAD+ | mechanism of dehydrogenation, mechanism of electron transfer: L-dihydroorotate transfers a pair of electrons to FMN via iron-sulfur cluster via FAD to NAD+ | Lactococcus lactis | orotate + NADH + H+ | - |
? | |
(S)-dihydroorotate + NAD+ | mechanism of dehydrogenation, mechanism of electron transfer: L-dihydroorotate transfers a pair of electrons to FMN via iron-sulfur cluster via FAD to NAD+ | Lactococcus lactis | orotate + NADH + H+ | - |
r | |
(S)-dihydroorotate + NAD+ | NAD+ as ultimate electron acceptor | Bacillus subtilis | orotate + NADH + H+ | - |
? | |
(S)-dihydroorotate + NAD+ | NAD+ as ultimate electron acceptor | Lactococcus lactis | orotate + NADH + H+ | - |
? | |
(S)-dihydroorotate + NAD+ | NAD+ as ultimate electron acceptor | Lactococcus lactis | orotate + NADH + H+ | - |
r | |
(S)-dihydroorotate + NAD+ | NAD+ as ultimate electron acceptor | Enterococcus faecalis | orotate + NADH + H+ | - |
? | |
(S)-dihydroorotate + NAD+ | NAD+ as ultimate electron acceptor | Faecalicatena orotica | orotate + NADH + H+ | - |
? | |
(S)-dihydroorotate + NAD+ | NAD+ as ultimate electron acceptor | Faecalicatena orotica | orotate + NADH + H+ | - |
r | |
dihydroorotate + NAD+ | fourth step and sole redox reaction in the pyrimidine de novo biosynthetic pathway | Lactococcus lactis | orotate + NADH + H+ | - |
r | |
dihydroorotate + NAD+ | fourth step and sole redox reaction in the pyrimidine de novo biosynthetic pathway | Enterococcus faecalis | orotate + NADH + H+ | - |
r | |
dihydroorotate + NAD+ | fourth step and sole redox reaction in the pyrimidine de novo biosynthetic pathway | Faecalicatena orotica | orotate + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
heterotetramer | alpha2,beta2 | Bacillus subtilis |
heterotetramer | alpha2,beta2, 2 * 32000 + 2 * 28000, SDS-PAGE | Faecalicatena orotica |
heterotetramer | alpha2,beta2, family IB enzyme | Lactococcus lactis |
heterotetramer | alpha subunit is named PyrDB, beta subunit is named PyrK | Bacillus subtilis |
heterotetramer | alpha subunit is named PyrDB, beta subunit is named PyrK | Lactococcus lactis |
Synonyms | Comment | Organism |
---|---|---|
More | family IB dihydroorotate dehydrogenase | Lactococcus lactis |
More | family IB dihydroorotate dehydrogenase | Faecalicatena orotica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Faecalicatena orotica |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.97 | - |
Orotate | formation of L-dihydroorotate, in absence of NAD+ or NADH, pH 8, 10 mM cysteine | Faecalicatena orotica | |
22 | - |
L-dihydroorotate | formation of orotate, in absence of NAD+ or NADH, pH 8, 10 mM cysteine | Faecalicatena orotica | |
45 | - |
L-dihydroorotate | in presence of NAD+ | Faecalicatena orotica |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.4 | 8.9 | below and above the turnover number decreases | Faecalicatena orotica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
4Fe-4S-center | iron-sulfur cluster resides on the beta subunit | Lactococcus lactis | |
4Fe-4S-center | cofactor content: 4 non-heme iron and 4 labile sulfur atoms per heterotetramer | Bacillus subtilis | |
4Fe-4S-center | cofactor content: 4 non-heme iron and 4 labile sulfur atoms per heterotetramer | Lactococcus lactis | |
4Fe-4S-center | cofactor content: 4 non-heme iron and 4 labile sulfur atoms per heterotetramer | Enterococcus faecalis | |
4Fe-4S-center | cofactor content: 4 non-heme iron and 4 labile sulfur atoms per heterotetramer | Faecalicatena orotica | |
FAD | FAD is located on the beta subunit | Lactococcus lactis | |
FAD | two molecules FAD per heterotetramer | Bacillus subtilis | |
FAD | two molecules FAD per heterotetramer | Lactococcus lactis | |
FAD | two molecules FAD per heterotetramer | Enterococcus faecalis | |
FAD | two molecules FAD per heterotetramer | Faecalicatena orotica | |
FAD | flavoprotein, ratio FAD to FMN 1:1 | Faecalicatena orotica | |
FMN | flavoprotein, ratio FAD to FMN 1:1 | Faecalicatena orotica | |
FMN | two molecules of FMN per heterotetramer | Bacillus subtilis | |
FMN | two molecules of FMN per heterotetramer | Lactococcus lactis | |
FMN | two molecules of FMN per heterotetramer | Enterococcus faecalis | |
FMN | two molecules of FMN per heterotetramer | Faecalicatena orotica | |
FMN | FMN is located on the alpha subunit | Lactococcus lactis |