Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.4.2.3 extracted from

  • Visser, D.; Hennessy, F.; Rashamuse, J.; Pletschke, B.; Brady, D.
    Stabilization of Escherichia coli uridine phosphorylase by evolution and immobilization (2011), J. Mol. Catal. B, 68, 279-285.
No PubMed abstract available

Application

Application Comment Organism
synthesis evolvement of a mutant enzyme by iterative saturation mutagenesis. Compared to the wild type enzyme, which has a temperature optimum of 40°C and a half-life of 9.89 h at 60°C, the selected mutant has a temperature optimum of 60°C and a half-life of 17.3 h at 60°C. Self-immobilization of the native enzyme as a Spherezyme shows a 3.3fold increase in thermostability while immobilized mutant enzyme shows a 4.4fold increase in thermostability. Combining the enzyme with the purine nucleoside phosphorylase from Bacillus halodurans allows for synthesis of 5-methyluridine (a pharmaceutical intermediate) from guanosine and thymine in a one-pot transglycosylation reaction. Replacing the wild type uridine phosphorylase with the mutant allows for an increase in reaction temperature to 65°C and increased the reaction productivity from 10 to 31 g per l and h Escherichia coli

Protein Variants

Protein Variants Comment Organism
additional information evolvement of a mutant enzyme UPL8 by iterative saturation mutagenesis. Compared to the wild type enzyme, which has a temperature optimum of 40°C and a half-life of 9.89 h at 60°C, the selected mutant has a temperature optimum of 60°C and a half-life of 17.3 h at 60°C. Self-immobilization of the native enzyme as a Spherezyme shows a 3.3fold increase in thermostability while immobilized mutant enzyme shows a 4.4fold increase in thermostability Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
wild-type Escherichia coli
60
-
mutant UPL8 Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
75
-
1 h, 3.7% residual activity for wild-type, 80.2% residual activity for mutant UPL8 Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
5.6 8.4 mutant UPL8, more than 60% of maximum activity within Escherichia coli
6 8.2 wild-type, more than 60% of maximum activity within Escherichia coli