Table 3. Thermal stability of wild-type and engineered adGSTD4-4 mutants at 45 °C.
The remaining GST activity was measured after incubating the enzyme at various time points at 45 °C.
| Enzyme | Half-life (t1/2) at 45 °C (min) |
|---|---|
| Wild-type | 15.3±0.31 |
| L33A | 45.3±1.56* |
| L33Y | 71.7±1.71* |
| L33F | 212±17.9* |
| L33I | 30.4±1.5* |
| L6A | 1.73±0.01* |
| T31A | 22.7±1.77* |
| I52A | 9.58±0.65* |
| I52L | 18.1±1.33 |
| E37A | 1.73±0.04* |
| E37Q | 6.54±0.54* |
| K40A | 3.94±0.20* |
| E42A | 8.44±0.48* |
| A35R | 9.01±0.54* |
ANOVA analysis revealed a significant difference compared with the wild-type enzyme, where indicated (*P<0.001); the absence of an asterisk indicates no significant difference compared with wild-type (in these experiments, exclusively the I52L mutant).