Table 2 |
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| Kinetic properties of PpAOS2 with different hydroperoxy fatty acid substrates | ||||
| Substrate | KM[μM] | Vmax[μM/min] | kcat[1/min] | kcat/KM[min-1M-1× 106] |
| 9-HPOD | 36 +/− 5 | 0.02 +/− 0.001 | 5 | 0.14 |
| 9- HPOT(n-3) | 40 +/− 4 | 0.01 +/− 0.001 | 2.5 | 0.06 |
| 9- HPOT(n-6) | 28 +/− 4. | 0.03 +/− 0.002 | 7.5 | 0.27 |
| 13-HPOD | 27+/− 3 | 0.04 +/− 0.002 | 10 | 0.37 |
| 13- HPOT(n-3) | 30 +/− 5 | 0.02 +/− 0.002 | 5 | 0.17 |
| 13- HPOT(n-6) | 42 +/− 10 | 0.02 +/− 0.002 | 5 | 0.12 |
| 12-HPETE | 10 +/− 5 | 0.49 +/− 0.057 | 12250 | 1228.69 |
Kinetic properties were determined by measuring the initial time-dependent substrate consumption at 234 nm at different substrate concentrations typically ranging from 2–100 μM. For analysis between 20 and 30 data points data points were fitted to the Michaelis-Menten equation. Note that the PpAOS2-concentration used for incubations with 12-HPETE was different (1 nM) from those used for incubations with the other substrates (100 nM). Kcat values are corrected to 100% heme occupancy from the ~4% heme content in the enzyme preparation.
Scholz et al. BMC Plant Biology 2012 12:228 doi:10.1186/1471-2229-12-228
