Table 2.
Apparent steady-state parameters and FMN contents of KefF
| KefF | FMN/KefF ratio (mol/mol) | kcatapp(NADH) (s−1) | Kmapp (NADH) (mM) | kcatapp (MTT) (s−1) | Kmapp (MTT) (mM) |
|---|---|---|---|---|---|
| WT (NADH) | 0.4 ± 0.1 | 1.9 ± 0.6 | 0.15 ± 0.04 | 2.2 ± 0.8 | 0.4 ± 0.1 |
| WT (NADPH)a | 0.4 ± 0.1 | 1.7 ± 0.5 | 0.07 ± 0.06 | 2.7 ± 1.9 | 0.7 ± 0.6 |
| G107S | not detectable | 0.04 ± 0.03 | 1.1 ± 0.1 | 0.015 ± 0.005 | 0.13 ± 0.06 |
| H112W | 0.61 ± 0.05 | 0.28 ± 0.04 | 4.7 ± 2.5 | 0.06 ± 0.03 | 0.02 ± 0.01 |
| F149W | 0.7 ± 0.1 | 0.3 ± 0.1 | 0.05 ± 0.02 | 0.3 ± 0.1 | 0.10 ± 0.05 |
| C147S | 0.5 ± 0.1 | 2.1 ± 0.2 | 0.06 ± 0.04 | 2.7 ± 0.6 | 0.43 ± 0.06 |
| C151S | 0.53 ± 0.07 | 2.6 ± 0.7 | 0.07 ± 0.03 | 3.2 ± 0.7 | 0.33 ± 0.06 |
| C147S/C151S | 0.42 ± 0.07 | 2.0 ± 0.7 | 0.05 ± 0.03 | 2.9 ± 0.9 | 0.53 ± 0.06 |
a
The values in this row are for the reaction of NADPH instead of NADH.