Table 1.
Construct | Quantum Yield | Specific Activitya | |
---|---|---|---|
FAD | Trp | ||
WT | 0.06 | 0.08 | 36±4 |
0.05b | 0.08 | ||
W47F | 0.05 | 0.05 | 48±3 |
0.05 | 0.05 | ||
W131F | 0.02 | 0.11 | 20±3 |
0.04 | 0.10 | ||
W52F | n.d.c | 0.12 | 42±10 |
n.d.c | 0.14 | ||
Y137F | n.d.c | 0.17 | 32±4 |
Specific activity (µmol NADH oxidized per minute per mg enzyme) was measured at 22–23 °C with 200 µM NADH and 120 µM FAD.
Duplicate entries represent experiments with independent preparations of NOX. The experimental errors of replicate measurements of the quantum yield with the same preparation were approximately ±0.002 for FAD and ±0.01 for Trp.
Flavin fluorescence for these constructs was too low to measure QFAD reliably.