TABLE 1.
Steady-state kinetic constants of native AAO and three Phe-501 variants for p-methoxybenzyl alcohol and O2 substrates
Maximal steady-state kinetic constants were determined by varying simultaneously the concentrations of p-methoxybenzyl alcohol (Al) and oxygen (Ox) in 0.1 m phosphate (pH 6) at 12 °C. Rate constants were fitted to Equations 1 (native) and 2 (directed variants). Means ± S.D. are provided.
| kcat | Km(Al) | kcat/Km(Al) | Km(Ox) | kcat/Km(Ox) | |
|---|---|---|---|---|---|
| s−1 | μm | s−1mm−1 | μm | s−1mm−1 | |
| AAO | 105 ± 1 | 29 ± 1 | 3620 ± 80 | 134 ± 3 | 784 ± 20 |
| F501Y | 87 ± 1 | 17 ± 1 | 5120 ± 310 | 180 ± 5 | 483 ± 13 |
| F501W | 64 ± 1 | 249 ± 5 | 257 ± 6.5 | 46 ± 2 | 1390 ± 50 |
| F501A | 40 ± 1 | 167 ± 5 | 240 ± 9 | 3600 ± 110 | 11 ± 0.5 |