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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
wild-type | Escherichia coli |
60 | - |
mutant UPL8 | Escherichia coli |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Escherichia coli |
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 |