Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 cells | [Bacillus] caldolyticus |
expressed in Escherichia coli SO1263 cells | Mesocricetus auratus |
expression in Escherichia coli strain SØ1263/pyrC- | Cricetinae |
overexpression in Escherichia coli BL21(DES/pLysS) | [Bacillus] caldolyticus |
Protein Variants | Comment | Organism |
---|---|---|
D230G | catalytically inactive | [Bacillus] caldolyticus |
D230N | catalytically inactive | [Bacillus] caldolyticus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid | at 37°C or 60°c the enzyme binds 2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid only slightly more strongly than DHO; competitive inhibitor | [Bacillus] caldolyticus | |
2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid | at 37°C or 60°C the enzyme binds 2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid more tightly than DHO | Cricetinae | |
2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid | competitive inhibitor | Mesocricetus auratus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | tagged enzyme contains ca. 1.0 zinc atom per subunit measured by atomic absorption spectroscopy | [Bacillus] caldolyticus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
48000 | - |
gel filtration | [Bacillus] caldolyticus |
48000 | - |
gel filtration, pure tagged enzyme | [Bacillus] caldolyticus |
48090 | - |
mass spectroscopy, BcDHOase with the hexahistidine tag | [Bacillus] caldolyticus |
48090 | - |
electrospray ionization mass spectroscopy, enzyme with hexahistidine tag | [Bacillus] caldolyticus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cricetinae | - |
- |
- |
Mesocricetus auratus | - |
- |
- |
[Bacillus] caldolyticus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
by chromatography on N-linked gutylamine-agarose and DEAE-Sephacel | Cricetinae |
by Ni-NTA chromatography and Poros Q anion-exchange column | [Bacillus] caldolyticus |
N-linked butylamine-agarose column chromatography and DEAE-Sephacel column chromatography | Mesocricetus auratus |
Ni-NTA agarose column chromatography, Poros Q column chromatography, and Sephadex G-75 gel filtration | [Bacillus] caldolyticus |
Storage Stability | Organism |
---|---|
-20°C, Na HEPES, pH 7.3, 10% (v/v) glycerol | Cricetinae |
4°C, Na HEPES, pH 7.5 | [Bacillus] caldolyticus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-dihydroorotate + H2O | - |
[Bacillus] caldolyticus | N-carbamoyl-L-aspartate | - |
r | |
L-dihydroorotate + H2O | - |
Cricetinae | N-carbamoyl-L-aspartate | - |
r | |
N-carbamoyl-L-aspartate | - |
Mesocricetus auratus | L-dihydroorotate + H2O | - |
r | |
N-carbamoyl-L-aspartate | - |
[Bacillus] caldolyticus | L-dihydroorotate + H2O | - |
r |
Subunits | Comment | Organism |
---|---|---|
monomer | - |
[Bacillus] caldolyticus |
Synonyms | Comment | Organism |
---|---|---|
BcDHOase | - |
[Bacillus] caldolyticus |
DHOase | - |
Mesocricetus auratus |
DHOase | - |
[Bacillus] caldolyticus |
hDHOase | - |
Cricetinae |
pyrC | - |
[Bacillus] caldolyticus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
- |
Mesocricetus auratus |
37 | - |
- |
Cricetinae |
70 | - |
- |
[Bacillus] caldolyticus |
70 | - |
BcDOHAse starts to unfold at temperatures greater than 70°C | [Bacillus] caldolyticus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
stable for 10 min | Cricetinae |
50 | - |
at 50°C the enzyme remains stable for 10 min | Mesocricetus auratus |
70 | - |
BcDHOase starts to unfold at temperatures greater than 70°C with a melting temperature of 79.5°C | [Bacillus] caldolyticus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0045 | - |
2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid | at 37°C | Mesocricetus auratus | |
0.083 | - |
2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid | at 37°C | [Bacillus] caldolyticus | |
0.102 | - |
2-oxo-1,2,3,6-tetrahydropyrimidine-4,6-dicarboxylic acid | at 60°C | [Bacillus] caldolyticus |