Table 1.
Characteristics of reconstructed and experimentally characterized ancestral haloalkane dehalogenases
| Protein code | Expression (% of total protein) | Solubility (%) | Yield (mg/l) | T m (°C) | HLD act. (μmol/mg·s) | LUC act. (RLU/mg·s) |
|---|---|---|---|---|---|---|
| DhaA wt | 17 | 83.1 | 91.1 | 50.56 ± 2.4 | 0.032 ± 0.0059 | n.a. |
| DhaA 172Loc | 23 | 85.5 | 74.9 | 71.60 ± 0.7 | 0.038 ± 0.0002 | 1.41 ± 0.26 |
| DhaA 172Glob | 21 | 65.2 | 88.2 | 70.04 ± 1.5 | 0.061 ± 0.0045 | n.a. |
| DhaA 230Loc | 20 | n.d. | n.d. | n.d. | n.d. | n.d. |
| DhaA 230Glob | 23 | 84.8 | 108.5 | 72.14 ± 0.4 | 0.061 ± 0.0118 | n.a. |
| DhaA 238Loc | 23 | 63.2 | 74.9 | 70.36 ± 0.6 | 0.014 ± 0.0021 | 353.5 ± 14.58 |
| DhaA 238Glob | 19 | 83.3 | 94.4 | 76.19 ± 0.2 | 0.030 ± 0.0012 | 3.18 ± 0.33 |
Notes: DhaA, haloalkane dehalogenase from Rhodococcus rhodochrous NCIMB 13064; wt, wild type; Loc, ancestral protein inferred from local alignment; Glob, ancestral protein inferred from global alignment; Tm, melting temperature; HLD act., haloalkane dehalogenases activity; LUC act., luciferase activity; n.d., not determined due to poor solubility of this protein; n.a., not active under tested conditions.