Table 2.
Substrate | Enzyme |
|
|
|
b |
|
|
---|---|---|---|---|---|---|---|
s−1 | μM | × 105 s−1 M−1 | s−1 | μM | × 105 s−1 M−1 | ||
lignostilbene | WT | 18 ± 1 | 13 ± 1 | 14 ± 1 | 39 ± 2 | 320 ± 20 | 1.2 ± 0.1 |
S283A | 15.3 ± 0.4 | 10 ± 1 | 15 ± 1 | 20 ± 1 | 400 ± 30 | 0.50 ± 0.04 | |
S283F | 0.14 ± 0.01 | 8.5 ± 0.7 | 0.16 ± 0.02 | n.d.c | n.d. | n.d. | |
DCA-S | WT | 11 ± 1 | 12 ± 1 | 9 ± 1 | 26 ± 1 | 220 ± 20 | 1.2 ± 0.1 |
S283A | 4.1 ± 0.1 | 4.6 ± 0.5 | 9 ± 1 | 7.8 ± 0.6 | 410 ± 60 | 0.19 ± 0.03 | |
S283F | 0.035 ± 0.001 | 2.7 ± 0.5 | 0.13 ± 0.05 | n.d. | n.d. | n.d. |
Experiments were performed using HEPES (I = 0.1 M), pH 7.5, at 25 °C. Parameters were calculated using a minimum of 20 data points at various substrate concentrations. Apparent parameters for stilbene substrates were obtained using air-saturated buffer. Parameters obtained for O2, including kcat values, were determined using 100 μM lignostilbene or DCA-S.
Calculated from saturating amounts of O2 and stilbene.
Not determined.