Table III.
Kinetic parameters with respect to the salutaridine of mutated variants of SalR designed for the investigations on catalytic site residues and substrate binding
NADPH concentration 250 μm.
| Variant | Km | Ki | Vopt | Kcat | KcatKm−1 |
|---|---|---|---|---|---|
| μm | μm | nkat mg−1 | s−1 | s−1m−1 103 | |
| Wild type | 7.9 ± 2.8 | 140 ± 57 | 63.2 ± 8 | 2.3 ± 0.3 | 291 |
| N152A | 7.9 ± 2.6 | 229 ± 81 | 9.8 ± 1.3 | 0.36 ± 0.05 | 46 |
| S180A | 25.3 ± 3.6 | 179 ± 19 | 0.068 ± 0.01 | 0.0025 ± 0.0002 | 0.099 |
| K240E | Inactive | ||||
| Y236F | Inactive | ||||
| V106A | 9.4 ± 1.1 | 168 ± 21 | 121 ± 0.9 | 4.5 ± 0.3 | 479 |
| F104A | 52.5 ± 6.2 | 203 ± 33 | 123 ± 10 | 4.5 ± 0.3 | 86 |
| L266A | 96 ± 6.9 | 840 ± 82 | 34.2 ± 1.6 | 1.3 ± 0.06 | 13 |
| L266S | 98.3 ± 6.3 | 1,356 ± 143 | 30.0 ± 1.1 | 1.1 ± 0.05 | 11 |
| L266V | 44.5 ± 3.8 | 1,423 ± 197 | 50.3 ± 2.3 | 1.8 ± 0.09 | 40 |
| M271T | Inactive | ||||
| N272T | Inactive | ||||