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Literature summary for 3.5.2.3 extracted from

  • Huang, D.T.; Kaplan, J.; Menz, R.I.; Katis, V.L.; Wake, R.G.; Zhao, F.; Wolfenden, R.; Christopherson, R.I.
    Thermodynamic analysis of catalysis by the dihydroorotases from hamster and Bacillus caldolyticus, as compared with the uncatalyzed reaction (2006), Biochemistry, 45, 8275-8283.
    View publication on PubMed
Search for more literature for 3.5.2.3

Cloned(Commentary)

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

Protein Variants Comment Organism
D230G catalytically inactive [Bacillus] caldolyticus
D230N catalytically inactive [Bacillus] caldolyticus

Inhibitors

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

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 [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

Organism UniProt Comment Textmining
Cricetinae
-
-
-
Mesocricetus auratus
-
-
-
[Bacillus] caldolyticus
-
-
-

Purification (Commentary)

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

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 and Products (Substrate)

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

Subunits Comment Organism
monomer
-
 [Bacillus] caldolyticus

Synonyms

Synonyms Comment Organism
BcDHOase
-
 [Bacillus] caldolyticus
DHOase
-
 Mesocricetus auratus
DHOase
-
 [Bacillus] caldolyticus
hDHOase
-
 Cricetinae
pyrC
-
 [Bacillus] caldolyticus

Temperature Optimum [°C]

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 [°C]

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 [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