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Literature summary extracted from

  • Del Arco, J.; Galindo, J.; Clemente-Suarez, V.J.; Corrales, A.; Fernandez-Lucas, J.
    Sustainable synthesis of uridine-5-monophosphate analogues by immobilized uracil phosphoribosyltransferase from Thermus thermophilus (2020), Biochim. Biophys. Acta, 1868, 140251 .
    View publication on PubMed

Application

EC Number Application Comment Organism
2.4.2.9 synthesis MtTtUPRT is successfully applied in the enzymatic synthesis of therapeutic NMP analogues. The immobilized enzyme is usable as a biocatalyst in the synthesis of different 5-modified uridine-5'-monophosphates at short times. MTtUPRT3 can be reused up to 8 times in the synthesis of uridine-5'-monophosphate (UMP) with maximum activity at 100°C and pH 7.0 (activity 968 IU/g support, retaining activity 100%). Potential application of MTtUPRT3 as industrial biocatalyst Thermus thermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.9 recombinant expression of N-terminal His6-tagged TtUPRT Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
2.4.2.9 additional information covalent immobilization of uracil phosphoribosyltransferase (UPRT) from Thermus thermophilus HB8 onto glutaraldehyde-activated magnetic iron oxide porous microparticles (MTtUPRT). The support is activated by contacting the microspheres with 0.2 ml of 50 mM sodium phosphate buffer, pH 8.5, containing 5% w/v glutaraldehyde during 3 h at 25°C, and then washed thoroughly with distilled water to remove the excess activating agent. Finally, activated microspheres are washed and equilibrated in binding buffer (50 mM potassium phosphate buffer, pH 8.5), shortly prior to use. According to the catalyst load experiments, MTtUPRT3 is selected as optimal biocatalyst. MTtUPRT3 is active and stable in a broad range of temperature (70-100°C) and in the pH interval 6.0-8.0, displaying maximum activity at 100°C and pH 7.0 (activity 968 IU/g support, retaining activity 100%). In addition, MTtUPRT3 can be reused up to 10 times in the synthesis of uridine-5'-monophosphate (UMP), completely stable for 6 cycles. Biocatalyst recycling is studied by employing MTtUPRT3 in consecutive reactions. MTtUPRT3 is successfully applied in the sustainable synthesis of different 5-modified uridine-5'-monophosphates at short times Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.9 Mg2+ required for catalysis Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.9 UMP + diphosphate Thermus thermophilus
-
 uracil + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.9 Thermus thermophilus Q5SIQ7
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.9 5-bromo-UMP + diphosphate
-
 Thermus thermophilus 5-bromouracil + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?
2.4.2.9 5-chloro-UMP + diphosphate
-
 Thermus thermophilus 5-chlorouracil + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?
2.4.2.9 5-fluoro-UMP + diphosphate
-
 Thermus thermophilus 5-fluorouracil + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?
2.4.2.9 5-iodo-UMP + diphosphate
-
 Thermus thermophilus 5-iodouracil + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?
2.4.2.9 UMP + diphosphate
-
 Thermus thermophilus uracil + 5-phospho-alpha-D-ribose 1-diphosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.2.9 homotetramer structure modeling, overview Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
2.4.2.9 TtUPRT
-
 Thermus thermophilus
2.4.2.9 TtUPRT3
-
 Thermus thermophilus
2.4.2.9 UPP
-
 Thermus thermophilus
2.4.2.9 UPRT
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.9 60
-
 free enzyme Thermus thermophilus
2.4.2.9 100
-
 immobilized enzyme Thermus thermophilus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.4.2.9 40 100 over 40% of maximal activity within this range, profile overview Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.9 7
-
 immobilized enzyme Thermus thermophilus
2.4.2.9 8
-
 free enzyme Thermus thermophilus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.2.9 4 10 over 60% of maximal activity within this range, profile overview Thermus thermophilus

General Information

EC Number General Information Comment Organism
2.4.2.9 physiological function uracil phosphoribosyltransferase (UPRT) catalyzes the reversible transfer of the 5-phosphoribosyl group from 5-phospho-alpha-D-ribosyl1-diphosphate (PRPP) to uracil N1 to form uridine-5'-monophosphate, UMP, in the presence of Mg2+. Nucleosides are essential metabolites in numerous biochemical processes in all living organisms Thermus thermophilus