Table I.
Kinetic behavior of wt-CocE and mutant CocE
| CocE variant | τ1/2 (min) | Kcat (s−1) | Km (mM) | Kcat/Km (s−1 M−1) |
|---|---|---|---|---|
| G173Q | 75.0 | 36.1 ± 1.11 | 0.0037 ± 0.0017 | 9.88 × 106 |
| L169K | 570 | 80.1 ± 10.8 | 0.044 ± 0.013 | 1.84 × 106 |
| T172R | 78.0 | 50.9 ± 15.9 | 0.017 ± 0.0047 | 3.06 × 106 |
| T172R/G13Q | 370 | 53.4 ± 0.65 | 0.017 ± 0.004 | 3.18 × 106 |
| T172R/G173Q/L169K | NA | No fit | No fit | NA |
| wt-CocE | 12.2 | 51.4 ± 16.8 | 0.0057 ± 0.0019 | 8.99 × 106 |
The metabolism of cocaine by purified preparations of wt-CocE, T172R, G173Q, T172R/G173Q or L169K CocE was measured as described in Materials and Methods section.
NA, not applicable; No fit, no Michaelis–Menton kinetics.