TABLE 2.
Calculated kinetic parameters of Rv2361ca
| Parameter | Substrate
|
|||||
|---|---|---|---|---|---|---|
| IPP | GPP | NPP | E,E-FPP | E,Z-FPP | GGPP | |
| Km (μM) | 89 | 490 | 29 | 84 | 290 | 40 |
| Vmax (pmol/min) | 34 | 25 | 21 | 30 | 4,800 | 12 |
| Kcat (min−1) | 4.8 | 3.6 | 3.1 | 4.3 | 690 | 1.8 |
| Kcat/Km | 5.4 × 104 | 7.4 × 103 | 1.1 × 105 | 5.1 × 104 | 2.4 × 106 | 4.3 × 105 |
a
To determine the kinetic parameters, assays were done in the presence of 30 μM [14C]IPP and various concentrations of an allylic primer or of 100 μM ω,E,E-FPP and various concentrations of [14C]IPP. Reaction conditions were as described in Materials and Methods, and 6.9 pmols of Rv2361c was used to initiate the reaction. The data were subjected to nonlinear regression analysis using SigmaPlot for Windows.