TABLE 5.
Kinetic constants of wild-type PhaJAc and mutants L65A and V130G at 30°C
| Substrate | Wild-type enzyme
|
L65A
|
V130G
|
||||||
|---|---|---|---|---|---|---|---|---|---|
| Km (μM) | kcat (s−1) | kcat/Km (s−1 M−1) | Km (μM) | kcat (s−1) | kcat/Km (s−1 M−1) | Km (μM) | kcat (s−1) | kcat/Km (s−1 M−1) | |
| Crotonyl-CoA (C4) | 24 ± 2 | 1,922 ± 94 | 8.0 × 107 | 33 ± 3 | 899 ± 51 | 2.7 × 107 | 154 ± 11 | 3,141 ± 127 | 2.0 × 107 |
| Hexenoyl-CoA (C6) | 40 ± 5 | 294 ± 23 | 7.4 × 106 | 18 ± 2 | 89 ± 3 | 4.9 × 106 | 102 ± 12 | 402 ± 25 | 3.9 × 106 |
| Octenoyl-CoA (C8) | 42 ± 5 | 0.58 ± 0.04 | 1.4 × 104 | 21 ± 2 | 27 ± 1 | 1.3 × 106 | 76 ± 6 | 405 ± 14 | 5.3 × 106 |
| Decenoyl-CoA (C10) | 42 ± 4 | 0.65 ± 0.04 | 1.5 × 104 | 27 ± 4 | 18 ± 1 | 6.7 × 105 | 13 ± 2 | 108 ± 5 | 8.3 × 106 |
| Dodecenoyl-CoA (C12) | 43 ± 5 | 0.15 ± 0.01 | 3.5 × 103 | 34 ± 3 | 1.7 ± 0.1 | 5.0 × 104 | 5 ± 1 | 16 ± 1 | 3.2 × 106 |