Table 3. Characteristics of predicted multiple-point mutants of LinA.
| Method | Protein | Mutations | Rosetta ΔΔG (kcal.mol-1) | DSC | Activity at 30°C b | ||
|---|---|---|---|---|---|---|---|
| T m (°C) | ΔT m (°C) | (μmol s-1 mg-1) | |||||
| LinAwt | - a | - a | 41.4 ± 0.1 | - a | 0.21 (0.12 mM) c | 1.91 (0.38 mM) c | |
| Energy-based | LinA01 | D3I + S127Y + T133I + A145H | -31.4 | 62.3 ± 0.4 | +20.9 | 0.32 (0.12 mM) c | 1.29 (0.34 mM) c |
| Evolution-based | LinA02 | Y50F + F68W + A131V | 0.1 | 37.7 ± 0.2 | -3.7 | 0.17 (0.11 mM) c | ND |
a not applicable
b activity determined with γ-hexachlorocyclohexane at 30°C and pH 8.6
c initial γ-HCH concentration is given since it affects determined specific activity; ΔΔG–predicted change in Free Gibbs Energy; DSC–Differential Scanning Calorimetry; ND, not determined