Table 2.
Alcohol and NADPH oxidation kinetics of the fusion enzymes.[a]
|
Enzyme |
Cyclohexanol oxidation |
NADPH oxidation |
||||
|---|---|---|---|---|---|---|
|
|
k cat |
K M |
k cat /K M |
k cat |
K M |
k cat /K M |
|
|
[s−1] |
[mm] |
[s−1 m −1] |
[s−1] |
[μm] |
[s−1 mm −1] |
|
NOX |
– |
– |
– |
5.0[b] |
3.5[b] |
1400 |
|
ADHA |
0.26 |
19 |
14 |
– |
– |
– |
|
NOX‐A |
0.56 |
10 |
56 |
5.1 |
27 |
190 |
|
LbADH |
2.2 |
31 |
71 |
– |
– |
– |
|
NOX‐L |
2.0 |
29 |
69 |
4.4 |
5.8 |
760 |
|
TbADH |
8.3 |
3.7 |
2200 |
– |
– |
– |
|
T‐NOX |
5.7 |
5.8 |
980 |
2.8 |
5.7 |
490 |
[a] The kinetics were determined by measuring the change in absorbance at 340 nm at various concentrations of substrate (5–10 different concentrations, in duplicate or triplicate, Figures S4 and S5). The alcohol oxidation rates were measured in 20 mm KPO4 (pH 7.5), and NADPH oxidation was measured in 50 mm Tris⋅HCl (pH 8.0), both at 25 °C. [b] Data taken from ref. 9 (in 50 mm Tris⋅HCl (pH 7.5)).