Table 2.
Steady-state kinetic parameters of the enzymatic reactions catalyzed by TvFDPF1–3 and holoTvFDPF3
| Protein | Substrate | Km (μM) | Vmax (μmol min−1 mg−1) | kcat (s−1) | kcat/Km (s−1 M−1) |
|---|---|---|---|---|---|
| TvFDPF1 | NADH | 2.9 ± 0.8 | 4.9 ± 1.2 | 7.8 ± 1.9 | (2.6 ± 0.9) × 106 |
| NADPH | 1475 ± 486 | 4.9 ± 2.1 | 7.8 ± 3.3 | (5.2 ± 2.8) × 103 | |
| TvFDPF2 | NADH | 18 ± 2 | 6.1 ± 0.2 | 9.6 ± 0.3 | (5.3 ± 0.6) × 105 |
| NADPH | 1171 ± 208 | 1.1 ± 0.2 | 1.7 ± 0.3 | (1.4 ± 0.3) × 103 | |
| TvFDPF3 | NADH | 40 ± 8 | 12 ± 1 | 19 ± 2 | (4.8 ± 1.0) × 105 |
| NADPH | 399 ± 8 | 4.0 ± 0.1 | 6.4 ± 0.2 | (1.6 ± 0.0) × 104 | |
| holoTvFDPF3 | NADH | 56 ± 2 | 291 ± 26 | 466 ± 42 | (8.3 ± 0.8) × 106 |
| NADPH | 427 ± 160 | 11 ± 4 | 18 ± 6 | (4.1 ± 2.1) × 104 |
Activities were measured as described under the “Experimental procedures” section. Kinetic parameters represent the average of (n = 4–8) independent experiments ± SD. kcat values were calculated per monomer.