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Literature summary extracted from

  • Tiwari, K.; Kumar, R.; Dubey, V.K.
    Biochemical characterization of dihydroorotase of Leishmania donovani understanding pyrimidine metabolism through its inhibition (2016), Biochimie, 131, 45-53 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.2.3 gene DHO, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha, quantitative real-time PCR expression analysis Leishmania donovani

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.2.3 biotin sulfone
-
 Leishmania donovani
3.5.2.3 kaempferol kaempferol mediates perturbation of the pyrimidine pathway Leishmania donovani
3.5.2.3 additional information inhibitor docking study, overview Leishmania donovani

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.2.3 additional information
-
 additional information Michaelis-Menten kinetics Leishmania donovani
3.5.2.3 0.0281
-
 N-carbamoyl-L-aspartate pH 6.0, 25°C, recombinant enzyme Leishmania donovani
3.5.2.3 0.6024
-
 (S)-dihydroorotate pH 8.0, 25°C, recombinant enzyme Leishmania donovani

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.2.3 Zn2+ a zinc metalloenzyme Leishmania donovani

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.2.3 (S)-dihydroorotate + H2O Leishmania donovani
-
 N-carbamoyl-L-aspartate
-
 r
3.5.2.3 (S)-dihydroorotate + H2O Leishmania donovani BHU 1081
-
 N-carbamoyl-L-aspartate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
3.5.2.3 Leishmania donovani H9D0Z2
-
-
3.5.2.3 Leishmania donovani BHU 1081 H9D0Z2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.2.3 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Leishmania donovani

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.5.2.3 promastigote
-
 Leishmania donovani
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.3 (S)-dihydroorotate + H2O
-
 Leishmania donovani N-carbamoyl-L-aspartate
-
 r
3.5.2.3 (S)-dihydroorotate + H2O
-
 Leishmania donovani BHU 1081 N-carbamoyl-L-aspartate
-
 r
3.5.2.3 N-carbamoyl-L-aspartate preferred reaction direction Leishmania donovani (S)-dihydroorotate + H2O
-
 r
3.5.2.3 N-carbamoyl-L-aspartate preferred reaction direction Leishmania donovani BHU 1081 (S)-dihydroorotate + H2O
-
 r

Subunits

EC Number Subunits Comment Organism
3.5.2.3 ? x * 43900, about, sequence calculation, x* 48000, recombinant His6-tagged enzyme, SDS-PAGE Leishmania donovani

Synonyms

EC Number Synonyms Comment Organism
3.5.2.3 amidohydrolase family protein UniProt Leishmania donovani
3.5.2.3 DHO
-
 Leishmania donovani
3.5.2.3 LdDHOase
-
 Leishmania donovani

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.2.3 25
-
 assay at Leishmania donovani

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.2.3 1.1
-
 (S)-dihydroorotate pH 8.0, 25°C, recombinant enzyme Leishmania donovani
3.5.2.3 2.1
-
 N-carbamoyl-L-aspartate pH 6.0, 25°C, recombinant enzyme Leishmania donovani

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.2.3 6
-
 (S)-dihydroorotate formation Leishmania donovani
3.5.2.3 8
-
 (S)-dihydroorotate hydrolysis Leishmania donovani

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.5.2.3 0.0554
-
 biotin sulfone pH 8.0, 25°C, recombinant enzyme Leishmania donovani
3.5.2.3 0.1514
-
 kaempferol pH 8.0, 25°C, recombinant enzyme Leishmania donovani

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.2.3 Leishmania donovani sequence calculation
-
 6

General Information

EC Number General Information Comment Organism
3.5.2.3 evolution the enzyme belongs to the amidohydrolase superfamily Leishmania donovani
3.5.2.3 malfunction a parallel salvage pathway of pyrimidine biosynthesis exists, since kaempferol cannot completely inhibit the pathway in vivo Leishmania donovani
3.5.2.3 metabolism de novo pyrimidine biosynthesis pathway is well developed and functional in protozoan parasite Leishmania donovani. The dihydroorotase (LdDHOase) is the third enzyme of the pathway Leishmania donovani
3.5.2.3 additional information homology modeling of LdDHOase showing the active site residues, overview Leishmania donovani
3.5.2.3 physiological function the enzyme catalyzes the reversible cyclization of N-carbamyl aspartate to dihydroorotate Leishmania donovani

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.2.3 1.83
-
 (S)-dihydroorotate pH 8.0, 25°C, recombinant enzyme Leishmania donovani
3.5.2.3 74.7
-
 N-carbamoyl-L-aspartate pH 6.0, 25°C, recombinant enzyme Leishmania donovani