Table 3.
Kinetic properties of computationally redesigned epoxide hydrolases.
|
Variant |
Designed to produce |
Assay substrate |
ee [%][a] |
Preference |
k cat [s−1][b] |
K M [mm] [b] |
k cat/K M [M−1 s−1] |
|---|---|---|---|---|---|---|---|
|
LEH‐P |
– |
1 a |
13 |
(R,R) |
0.035±0.004 |
4.2±0.2 |
7.9 |
|
RR8 |
(R,R)‐1 b |
1 a |
85 |
(R,R) |
0.039±0.009 |
189±20 |
0.20 |
|
46C |
(R,R)‐1 b |
1 a |
34 |
(R,R) |
0.022±0.002 |
344±17 |
0.06 |
|
43A |
(S,S)‐1 b |
1 a |
8 |
(R,R) |
0.017±0.005[c] |
35±4[c] |
0.48 |
|
SS16 |
(S,S)‐1 b |
1 a |
−90 |
(S,S) |
0.062±0.001 |
54±4 |
1.14 |
|
24A |
(S,S)‐1 b |
1 a |
−85 |
(S,S) |
0.005±0.001 |
3±0.2 |
1.9 |
|
LEH‐P |
– |
2 a |
24 |
(R,R) |
0.056±0.010 |
18±1 |
3.1 |
|
32A |
(S,S)‐2 b |
2 a |
−82 |
(S,S) |
0.063±0.002 |
225±9 |
0.28 |
|
LEH‐P |
– |
3 a |
92 |
(R,R) |
0.147±0.012 |
0.37±0.02 |
406 |
|
60A |
(R,R)‐3 b |
3 a |
>99 |
(R,R) |
0.052±0.006 |
0.06±0.01 |
890 |
|
52A |
(R,R)‐3 b |
3 a |
−89 |
(S,S) |
0.003±0.001 |
0.16±0.03 |
19 |
|
41B |
(S,S)‐3 b |
3 a |
−94 |
(S,S) |
0.002±0.001 |
0.19±0.02 |
10.5 |
[a] Calculated from multiple data points that is, product enantiomers in different reactions. [b] Averages of duplicate measurements with standard deviation. [c] Single measurement; margins from average coefficient of variation for 1 a data.