Table 2.
Steady state kinetic parameters for wild-type CT and variants
| Construct | Vmax (min−1) | Km (biocytin, mM) | Km (malonyl-CoA, µM) |
|---|---|---|---|
| Wild-type CT | 211 ± 7 | 11.4 ± 0.4 | 40.0 ± 5.6 |
| C27A | 22.8 ± 1.4 | 11.5 ± 0.5 | 53.2 ± 9.2 |
| C30A | 1.5 ± 0.2 | 5.0 ± 1.1 | 20.2 ± 2.4 |
| C46A | 18.8 ± 0.7 | 4.0 ± 0.7 | 72.2 ± 10.4 |
| C49A | 26.7 ± 0.9 | 9.7 ± 0.6 | 106.8 ± 9.8 |
| C27/46A ‘double’ | 0 | ||
| C27/46/49A ‘triple’ | 0 | ||
| C27/30/46/49A ‘quadruple’ | 0 |
The apparent maximal velocities and Michaelis constant for biocytin were obtained by varying biocytin while holding malonyl-CoA constant at 100 µM. The apparent Michaelis constant for malonyl-CoA was obtained by varying malonyl-CoA while holding biocytin constant at 20 mM. The standard errors on Vmax and Km were determined from the nonlinear regression analysis.