Table 4.
Conversions of cyclohexanol and racemic 1‐phenylethanol by ADH/NOX fusion enzymes.
|
Substrate |
Enzyme |
Conversion |
ee [a] |
TTN |
TTN |
|---|---|---|---|---|---|
|
|
|
[%] |
[%] |
(enzyme) |
(cofactor) |
|
cyclohexanol |
NOX‐A |
95 |
n.a. |
31 666 |
475 |
|
|
NOX‐L |
99 |
n.a. |
33 000 |
495 |
|
|
T‐NOX |
69 |
n.a. |
23 000 |
345 |
|
rac‐1‐phenylethanol |
NOX‐A |
94 |
99 (R) |
31 333 |
470 |
|
|
NOX‐A |
56[b] |
99 (R) |
28 000 |
140 |
|
|
NOX‐L |
50 |
99 (S) |
16 666 |
250 |
Reaction conditions: 50 mm substrate with 1.5 μm of fusion enzyme in 50 mm Tris⋅HCl (pH 8.5), 100 μm NADP+, 64 h at 24 °C, 500 rpm (ThermoMixer Eppendorf); 10 μm FAD, 1000 U catalase. Reactions with rac‐1‐phenylethanol included 2 % DMSO. [a] Enantiomeric excess of the remaining alcohol substrate (Figure S6), [b] 0.5 μm NOX‐A, 25 mm substrate, 100 μm NADP+, 50 mm N‐cyclohexyl‐2‐aminoethanesulfonic acid (CHES; pH 9.0) for 24 h at 24 °C.