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Literature summary extracted from

  • Donini, S.; Ferraris, D.M.; Miggiano, R.; Massarotti, A.; Rizzi, M.
    Structural investigations on orotate phosphoribosyltransferase from Mycobacterium tuberculosis, a key enzyme of the de novo pyrimidine biosynthesis (2017), Sci. Rep., 7, 1180 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.10 expressed in Escherichia coli BL21(DE3) cells Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.2.10 hanging drop vapor diffusion method. Enzyme in complex with phosphate: 0.1 M HEPES pH 7.5, 10% (w/v) PEG8000, 8% (v/v) ethylene glycol. Enzyme in complex with 5-phospho-alpha-D-ribose 1-diphosphate: 0.1 M HEPES pH 7.5, 10% (w/v) PEG6000, 5% (v/v) 2-methyl-2,4-pentanediol. Enzyme in complex with ferric dicitrate: 0.01 M iron(III) chloride hexahydrate, 0.1 M sodium citrate tribasic dehydrate pH 6.1, 10% (v/v) and Jeffamine M-600 Mycobacterium tuberculosis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.10 Mg2+ required for activity, 20 mM used in assay conditions Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.10 orotate + 5-phospho-alpha-D-ribose 1-diphosphate Mycobacterium tuberculosis
-
orotidine 5'-phosphate + diphosphate
-
r
2.4.2.10 orotate + 5-phospho-alpha-D-ribose 1-diphosphate Mycobacterium tuberculosis H37Rv
-
orotidine 5'-phosphate + diphosphate
-
r
2.4.2.10 orotidine 5'-phosphate + diphosphate Mycobacterium tuberculosis
-
orotate + 5-phospho-alpha-D-ribose 1-diphosphate
-
r
2.4.2.10 orotidine 5'-phosphate + diphosphate Mycobacterium tuberculosis H37Rv
-
orotate + 5-phospho-alpha-D-ribose 1-diphosphate
-
r

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.10 Mycobacterium tuberculosis P9WHK9
-
-
2.4.2.10 Mycobacterium tuberculosis H37Rv P9WHK9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.2.10 Ni-NTA column chromatography and Sephacryl 200 gel filtration Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.2.10 2
-
crude enzyme, at pH 8.0, temperature not specified in the publication Mycobacterium tuberculosis
2.4.2.10 17
-
after 8.5fold purification, at pH 8.0, temperature not specified in the publication Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.10 orotate + 5-phospho-alpha-D-ribose 1-diphosphate
-
Mycobacterium tuberculosis orotidine 5'-phosphate + diphosphate
-
r
2.4.2.10 orotate + 5-phospho-alpha-D-ribose 1-diphosphate
-
Mycobacterium tuberculosis H37Rv orotidine 5'-phosphate + diphosphate
-
r
2.4.2.10 orotidine 5'-phosphate + diphosphate
-
Mycobacterium tuberculosis orotate + 5-phospho-alpha-D-ribose 1-diphosphate
-
r
2.4.2.10 orotidine 5'-phosphate + diphosphate
-
Mycobacterium tuberculosis H37Rv orotate + 5-phospho-alpha-D-ribose 1-diphosphate
-
r

Synonyms

EC Number Synonyms Comment Organism
2.4.2.10 OPRT
-
Mycobacterium tuberculosis
2.4.2.10 PyrE
-
Mycobacterium tuberculosis
2.4.2.10 rv0382c
-
Mycobacterium tuberculosis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.2.10 52
-
melting temperature Mycobacterium tuberculosis