TABLE 3.
Apparent kinetic parameters of the R. jostii Ro05698 Δ4-(5α)-KSTD wild type and active site mutants using 1-(5α)-AD as substrate
| Vmax | Km | kcat | kcat/Km | |
|---|---|---|---|---|
| nmol·mg−1·min−1 | μm | s−1 | s−1·m−1 | |
| Wild typea | 27 × 102 ± 1.5 × 102 | 10 ± 2 | 2.4 | 2.4 × 106 |
| W136A | NDb | ND | ND | ND |
| W136F | 8 × 102 ± 3 × 102 | 66 ± 25 | 0.7 | 1 × 104 |
| Y319F | ND | ND | ND | ND |
| Y466A | ND | ND | ND | ND |
| Y466F | ND | ND | ND | ND |
| S468T | 19 × 102 ± 3.6 × 102 | 17 × 101± 3.3 × 101 | 1.6 | 9 × 103 |
| S468A | ND | ND | ND | ND |
a Contains mutations A60T (nucleotide G178A) and T160A (nucleotide A478G), but these mutations do not or hardly have an effect on activity.
b ND, not detectable.