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The wild-type and mutant enzymes were incubated at 45 °C at the protein concentration of 1 mg/ml. The inactivation time courses were determined by taking suitable aliquots at the different time points for assay of remaining activity to calculate half-life of the enzyme.
| Enzyme | Half-life at 45 °C (min−1) |
|---|---|
| Wild-type | 2.33±0.12 |
| I52A | 0.42±0.01 |
| S65A | 1.70±0.08 |
| R66A | 145.73±10.47 |
| M101A | 5.72±0.71 |
