TABLE 2.
Steady-state kinetic parameters of acyl-CoA dehydrogenasesa
| Substrate | Enzyme | Km (μM) | kcat (s−1) | kcat/Km (M−1 s−1) |
|---|---|---|---|---|
| Cholyl-CoA | FadE34 | 47 ± 5.8 | 0.22 ± 0.0095 | (4.8 ± 0.20) × 103 |
| CasC | 5.7 ± 0.69 | 16 ± 0.61 | (2.8 ± 0.11) × 106 | |
| Deoxycholyl-CoA | FadE34 | 27 ± 3.5 | 0.44 ± 0.019 | (1.6 ± 0.069) × 104 |
| CasC | 2.2 ± 0.35 | 9.9 ± 0.60 | (4.4 ± 0.23) × 106 | |
| 3β-Hydroxy-5-cholen-24-oyl-CoA | FadE34 | 2.5 ± 0.79 | 3.7 ± 0.40 | (1.5 ± 0.16) × 106 |
| CasC | 3.0 ± 0.76 | 10 ± 1.0 | (3.5 ± 0.34) × 106 |
a
Acyl-CoA dehydrogenase assays were performed at 25°C and included 250 μM ferrocenium hexafluorophosphate in 100 mM TAPS buffer (pH 8.5).