Literature summary for 2.4.2.10 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 .

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	P9WHK9	-	-
Mycobacterium tuberculosis H37Rv	P9WHK9	-	-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms	Comment	Organism
OPRT	-	Mycobacterium tuberculosis
PyrE	-	Mycobacterium tuberculosis
rv0382c	-	Mycobacterium tuberculosis

Temperature Stability [°C]

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