Table 1. Observed Rates of FMN Reduction at 25 °C for PETNR Variants with NADPH and NADHa.
| kred (s–1) | KS (μM) | kred/KS (s–1 μM) | |
|---|---|---|---|
| NADPH | |||
| WT | 33.43 ± 0.22 | 103 ± 4 | (3.25 ± 0.13)×10–1 |
| L25A | 55.75 ± 0.35 | 199 ± 6 | (2.80 ± 0.09)×10–1 |
| L25I | 29.49 ± 0.49 | 130 ± 8 | (2.27 ± 0.14)×10–1 |
| I107A | 21.22 ± 0.25 | 130 ± 9 | (1.63 ± 0.11)×10–1 |
| I107L | 28.37 ± 0.20 | 107 ± 5 | (2.65 ± 0.12)×10–1 |
| NADH | |||
| WT | 2.03 ± 0.01 | 1067 ± 13 | (1.90 ± 0.03)×10–3 |
| L25A | 3.62 ± 0.04 | 2294 ± 91 | (1.58 ± 0.06)×10–3 |
| L25I | 1.74 ± 0.04 | 306 ± 40 | (5.69 ± 0.94)×10–3 |
| I107A | 2.05 ± 0.04 | 329 ± 37 | (6.23 ± 0.71)×10–3 |
| I107L | 1.94 ± 0.02 | 889 ± 36 | (2.18 ± 0.09)×10–3 |
a
Calculated from fitting data presented in Figure S5 to kobs1 = kred[NAD(P)H]/(KS + [NAD(P)H]). Reaction conditions: 50 mM potassium phosphate buffer (pH 7.0), 20 μM enzyme concentration, [NAD(P)H] ranging 0.1–50 mM, 25 °C.