BglB Variant Characterization Data
Hover over any column for a desciption of that parameter.
Click on any column header to sort the data by that value.
► Expand all averages to see data from individual experiments. ◄
☑ Rosetta/Foldit sequence numbering.
☐ Include noncurated data.
656 records found
Click on any column header to sort the data by that value.
► Expand all averages to see data from individual experiments. ◄
☑ Rosetta/Foldit sequence numbering.
☐ Include noncurated data.
656 records found
View complete BglB sequence.
For kinetic constants,
the table is color-coded by relative log values of
1/KM, kcat, and kcat/KM compared to WT.
log 1/KM is used so that larger values are "better".
For T50 and TM values and Rosetta scores, a linear scale is used.
Blue shading indicates a variant that is "worse" than WT.
Orange shading indicates a variant that is "better" than WT.
Variants shaded black expressed (as confirmed by gel electrophoresis and/or yield > 0.1 mg/mL).
Variants marked with an asterisk (*) expressed, but no yield was recorded.
How were these data calculated?
| relative scale | −5 | −4 | −3 | −2 | −1 | 0 | 1 |
|---|
the table is color-coded by relative log values of
1/KM, kcat, and kcat/KM compared to WT.
log 1/KM is used so that larger values are "better".
For T50 and TM values and Rosetta scores, a linear scale is used.
Blue shading indicates a variant that is "worse" than WT.
Orange shading indicates a variant that is "better" than WT.
Variants shaded black expressed (as confirmed by gel electrophoresis and/or yield > 0.1 mg/mL).
Variants marked with an asterisk (*) expressed, but no yield was recorded.
How were these data calculated?
| Variant ▲ |
Yield, cE (mg/mL) |
KM (mm) |
kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) |
Rosetta score change |
||
|---|---|---|---|---|---|---|---|---|---|
| ► | WT | 2.26 ± 1.69 | 4.95 ± 0.86 | 458.9 ± 23.5 | 121.82 ± 30.93 | 39.8 ± 0.7 | 44.9 ± 0.4 | — | |
| W11F | 0.71 | 5.33 ± 0.40 | 836.9 ± 16.4 | 156.97 ± 12.12 | — | — | — | ||
| G12N | — | — | — | — | — | — | — | ||
| ► | T13K | 0.69 | 0.95 ± 0.39 | 0.3 ± 0.0 | 0.36 ± 0.15 | — | 46.2 ± 2.1 | 36.0 | |
| ► | T13S | 0.92 ± 0.51 | 8.59 ± 0.93 | 312.0 ± 7.1 | 94.16 ± 8.85 | 39.0 ± 0.6 | 44.6 ± 0.3 | 2.8 | |
| ► | S14A | 1.69 | 5.57 ± 0.80 | 219.7 ± 7.9 | 46.68 ± 7.92 | 40.2 ± 0.3 | 43.8 ± 0.1 | 1.8 | |
| ► | T15A | * | 5.30 ± 0.48 | 504.4 ± 13.0 | 95.25 ± 8.88 | 38.2 ± 0.4 | 42.0 ± 0.0 | 9.3 | |
| S16A | * | 14.01 ± 0.40 | 125.6 ± 0.0 | 8.88 ± 0.22 | 39.4 ± 0.3 | 43.9 ± 0.0 | — | ||
| S16N | — | — | — | — | — | — | — | ||
| ► | S17A | 3.96 | 12.48 ± 2.65 | 445.8 ± 44.5 | 33.95 ± 6.06 | 40.2 ± 694,236.0 | — | — | |
| S17E | * | 7.32 ± 0.38 | 526.1 ± 6.5 | 71.87 ± 3.90 | 39.5 ± 0.1 | — | — | ||
| Y18A | * | 31.55 ± 3.61 | 162.4 ± 6.5 | 5.20 ± 0.65 | 38.3 ± 0.2 | 40.6 ± 0.0 | — | ||
| ► | Q19A | 1.25 | 1.68 ± 1.14 | 0.1 ± 0.0 | 0.08 ± 0.05 | 35.2 ± 0.7 | — | -1.3 | |
| Q19C | * | — | — | — | — | — | — | ||
| Q19D | 0.57 | — | — | — | — | — | — | ||
| ► | Q19M | 0.43 | — | — | — | — | 46.4 ± 1.5 | -18.7 | |
| Q19P | — | — | — | — | — | — | — | ||
| Q19S | * | — | — | — | 36.8 ± 0.2 | 43.1 ± 0.0 | — | ||
| ► | Q19T | 1.54 ± 0.65 | 3.37 ± 0.64 | 120.1 ± 3.1 | 24.01 ± 2.42 | — | 42.8 ± 0.0 | -0.3 | |
| I20N | 0.31 | 19.41 ± 4.87 | 44.5 ± 4.0 | 2.29 ± 0.61 | — | — | — | ||
| E21C | — | — | — | — | — | — | 5.4 | ||
| E21D | 0.41 | 4.55 ± 0.53 | 32.0 ± 1.0 | 7.04 ± 0.86 | 37.5 ± 0.3 | 37.5 ± 0.3 | 4.4 | ||
| E21F | — | — | — | — | — | — | 32.3 | ||
| E21G | — | — | — | — | — | — | 8.0 | ||
| E21H | — | — | — | — | — | — | 22.3 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| E21I | — | — | — | — | — | — | 32.6 | ||
| E21K | — | — | — | — | — | — | 27.7 | ||
| E21N | — | — | — | — | — | — | 6.4 | ||
| E21R | — | — | — | — | — | — | 16.9 | ||
| E21T | — | — | — | — | — | — | 6.5 | ||
| E21W | — | — | — | — | — | — | 383.6 | ||
| E21Y | 4.45 | 1.52 ± 0.20 | 184.4 ± 5.4 | 120.95 ± 16.52 | 40.3 ± 0.3 | 40.3 ± 0.3 | 27.1 | ||
| ► | G23A | 1.36 ± 0.63 | 1.36 ± 0.27 | 444.2 ± 17.7 | 323.14 ± 62.67 | 40.1 ± 0.2 | 45.0 ± 0.0 | 1.8 | |
| ► | D25Q | 3.48 ± 2.87 | 3.45 ± 0.63 | 307.9 ± 13.3 | 87.18 ± 12.62 | 40.3 ± 0.1 | 45.5 ± 0.0 | — | |
| E26K | 0.62 | 6.81 ± 0.48 | 638.8 ± 12.9 | 93.81 ± 6.93 | 38.4 ± 0.3 | — | -0.9 | ||
| S32L | — | — | — | — | — | — | — | ||
| W34A | — | — | — | — | — | — | — | ||
| D35E | 2.00 | 2.72 ± 0.31 | 16.3 ± 0.4 | 5.97 ± 0.70 | — | — | 11.6 | ||
| ► | D35W | 0.40 ± 0.06 | 2.62 ± 0.82 | 7.5 ± 0.5 | 2.74 ± 0.71 | — | 46.8 ± 3.6 | 14.3 | |
| T36I | — | — | — | — | — | — | — | ||
| T36N | 5.49 | 6.57 ± 1.21 | 23.9 ± 1.2 | 3.63 ± 0.69 | — | 46.8 ± 0.1 | -0.2 | ||
| F37N | 0.26 | 4.88 ± 0.15 | 8.3 ± 0.1 | 1.70 ± 0.06 | — | — | — | ||
| ► | C38S | 1.05 ± 0.20 | 5.96 ± 0.78 | 806.6 ± 22.6 | 168.26 ± 20.44 | 41.1 ± 0.3 | 42.2 ± 0.2 | 0.5 | |
| I40T | 1.41 | 4.05 ± 0.21 | 265.2 ± 3.8 | 65.54 ± 3.50 | — | 43.3 ± 0.2 | 2.2 | ||
| P41L | * | — | — | — | — | — | — | ||
| G46C | 0.04 | 40.81 ± 25.63 | 4.4 ± 1.2 | 0.11 ± 0.07 | — | — | — | ||
| C49S | 0.85 | 6.63 ± 3.19 | 570.0 ± 74.3 | 86.02 ± 42.89 | 40.0 ± 0.0 | — | — | ||
| V52A | 2.06 | 5.30 ± 0.91 | 107.2 ± 5.0 | 20.22 ± 3.61 | — | 40.8 ± 0.2 | — | ||
| V52G | * | 8.25 ± 0.54 | 562.9 ± 10.8 | 68.19 ± 4.55 | 39.3 ± 0.1 | — | — | ||
| ► | F57Y | 2.07 ± 0.68 | 4.65 ± 0.50 | 173.2 ± 4.9 | 40.65 ± 4.78 | — | 45.4 ± 0.1 | -0.1 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| H58R | 1.22 | 5.64 ± 0.68 | 665.5 ± 21.2 | 118.04 ± 14.68 | — | — | — | ||
| ► | H59R | 2.55 ± 1.58 | 5.90 ± 0.52 | 302.3 ± 6.9 | 74.39 ± 6.46 | — | 49.8 ± 0.3 | -0.2 | |
| K61A | 1.55 | 3.79 ± 0.40 | 167.6 ± 4.4 | 44.28 ± 4.78 | 38.7 ± 0.4 | — | 1.3 | ||
| ► | K61C | 2.86 ± 0.01 | 1.75 ± 0.23 | 115.8 ± 3.3 | 67.22 ± 8.78 | 38.1 ± 0.3 | — | 2.4 | |
| K61D | 1.11 | 3.96 ± 0.39 | 116.3 ± 2.8 | 29.36 ± 2.96 | 38.3 ± 0.3 | — | 5.6 | ||
| K61E | 1.36 | 3.79 ± 0.28 | 141.6 ± 2.7 | 37.39 ± 2.87 | 38.8 ± 0.2 | — | 0.8 | ||
| ► | K61F | 2.58 ± 0.71 | 3.29 ± 0.25 | 70.9 ± 1.6 | 23.62 ± 2.25 | 37.4 ± 0.5 | — | 7.6 | |
| ► | K61G | 1.36 ± 0.45 | 3.02 ± 0.21 | 102.4 ± 2.0 | 26.48 ± 2.04 | 38.0 ± 0.5 | — | 4.6 | |
| K61H | 6.16 | 1.95 ± 0.19 | 164.0 ± 3.6 | 84.15 ± 8.28 | 38.2 ± 0.1 | — | 3.3 | ||
| K61I | 1.80 | 2.43 ± 0.19 | 104.7 ± 2.0 | 43.13 ± 3.46 | 38.4 ± 0.2 | — | 1.1 | ||
| K61L | 0.83 | 2.37 ± 0.30 | 60.7 ± 1.9 | 25.68 ± 3.33 | 40.6 ± 0.8 | — | 5.5 | ||
| K61M | 4.60 | 4.59 ± 0.53 | 200.1 ± 6.0 | 43.57 ± 5.17 | 38.0 ± 0.1 | — | 2.6 | ||
| ► | K61N | 3.50 ± 0.22 | 4.16 ± 0.41 | 156.9 ± 4.4 | 37.68 ± 4.19 | 35.6 ± 7.2 | — | 3.1 | |
| K61P | 1.49 | 2.90 ± 0.26 | 81.6 ± 1.8 | 28.19 ± 2.62 | 38.9 ± 0.3 | — | 11.7 | ||
| K61Q | 1.42 | 4.28 ± 1.38 | 137.0 ± 12.5 | 31.98 ± 10.68 | 39.3 ± 0.2 | — | 0.8 | ||
| K61R | 2.60 | 2.82 ± 0.25 | 109.8 ± 2.3 | 38.93 ± 3.50 | 39.2 ± 0.3 | — | 1.4 | ||
| ► | K61S | 2.24 ± 1.27 | 3.22 ± 0.56 | 199.4 ± 6.8 | 77.69 ± 7.58 | 38.5 ± 0.2 | 46.2 ± 0.3 | 1.4 | |
| K61W | 1.94 | 3.95 ± 0.43 | 74.2 ± 2.2 | 18.78 ± 2.13 | 38.2 ± 0.7 | — | 7.8 | ||
| K61Y | 0.47 | 1.86 ± 0.17 | 93.3 ± 1.9 | 50.18 ± 4.73 | 38.0 ± 0.5 | — | 7.7 | ||
| D63A | 0.01 | — | — | 0.33 ± 0.09 | — | — | 0.0 | ||
| D63C | 0.98 | 0.69 ± 1.31 | 0.0 ± 0.0 | 0.07 ± 0.14 | — | — | — | ||
| D63F | 0.77 | 4.13 ± 9.33 | 0.1 ± 0.0 | 0.01 ± 0.03 | — | — | — | ||
| ► | D63M | 0.36 ± 0.26 | 0.13 ± 0.81 | 0.1 ± 0.1 | 1.96 ± 14.10 | — | — | — | |
| D63N | 0.44 | 0.28 ± 0.43 | 0.2 ± 0.0 | 0.56 ± 0.87 | — | — | — | ||
| D63P | 0.28 | — | — | — | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| D63R | 0.13 | — | — | 0.02 ± 0.00 | — | — | — | ||
| D63S | 0.35 | 0.09 ± 0.16 | 0.1 ± 0.0 | 1.39 ± 2.61 | — | — | — | ||
| ► | K68H | 3.26 ± 0.47 | 4.02 ± 0.85 | 326.4 ± 14.3 | 82.25 ± 14.18 | — | 46.1 ± 0.8 | 2.4 | |
| F72A | * | 5.47 ± 0.28 | 502.2 ± 6.5 | 91.79 ± 4.76 | 38.6 ± 0.4 | — | — | ||
| ► | Y75F | 1.99 ± 1.88 | 3.50 ± 1.43 | 126.6 ± 16.0 | 20.90 ± 8.88 | 40.1 ± 1.7 | 46.4 ± 0.6 | 2.0 | |
| ► | Y75N | 0.15 ± 0.06 | 4.98 ± 0.47 | 364.4 ± 11.9 | 102.68 ± 19.80 | 38.0 ± 1.2 | — | 10.8 | |
| R76A | — | — | — | — | — | — | — | ||
| R76L | 0.54 | 43.36 ± 1.82 | 7.4 ± 0.1 | 0.17 ± 0.01 | — | — | — | ||
| R76Q | 0.70 | 6.62 ± 2.33 | 0.8 ± 0.1 | 0.13 ± 0.05 | — | — | -0.1 | ||
| F77D | 0.22 | — | — | — | — | — | — | ||
| F77W | 0.38 | 73.32 ± 94.87 | 20.0 ± 14.3 | 0.27 ± 0.40 | — | — | — | ||
| S78A | 3.12 | 20.08 ± 1.37 | 190.0 ± 4.9 | 9.46 ± 0.69 | — | 37.5 ± 0.1 | 1.9 | ||
| ► | V79I | 3.56 ± 0.08 | 3.21 ± 0.31 | 508.4 ± 11.5 | 158.50 ± 16.22 | 39.2 ± 0.2 | — | — | |
| V79T | 2.67 | 5.29 ± 2.61 | 239.4 ± 31.8 | 45.23 ± 23.12 | — | 46.7 ± 0.1 | 4.7 | ||
| ► | A80P | 0.34 ± 0.04 | 4.13 ± 0.40 | 14.3 ± 0.4 | 3.21 ± 1.37 | 34.9 ± 0.5 | 30.6 ± 10.1 | 190.9 | |
| ► | A80S | 2.12 ± 0.82 | 4.65 ± 0.59 | 697.1 ± 29.8 | 134.68 ± 17.29 | 40.6 ± 0.4 | 42.9 ± 0.4 | 2.5 | |
| ► | A88E | 2.47 ± 0.38 | 3.87 ± 0.60 | 317.7 ± 12.0 | 190.09 ± 62.25 | — | 45.7 ± 0.2 | -1.4 | |
| A88K | 0.79 | 4.46 ± 0.60 | 307.4 ± 10.5 | 68.93 ± 9.53 | — | — | — | ||
| I90V | 1.25 | 6.67 ± 0.28 | 790.7 ± 9.2 | 118.48 ± 5.19 | — | — | — | ||
| ► | I91A | 0.42 ± 0.31 | 7.44 ± 2.58 | 603.9 ± 21.6 | 109.73 ± 11.98 | 38.5 ± 304,961.7 | — | -2.8 | |
| I91C | 5.45 | 3.58 ± 0.15 | 19.6 ± 0.2 | 5.47 ± 0.24 | 38.1 ± 0.2 | — | — | ||
| I91D | 0.72 | 11.21 ± 0.69 | 89.6 ± 1.8 | 7.99 ± 0.51 | 37.4 ± 183,653.0 | — | — | ||
| I91E | * | 6.71 ± 0.79 | 692.7 ± 28.1 | 103.26 ± 12.77 | 39.7 ± 0.3 | — | — | ||
| I91G | 0.45 | 9.30 ± 3.31 | 93.7 ± 10.2 | 10.08 ± 3.75 | — | — | — | ||
| I91H | 3.69 | 11.59 ± 0.81 | 181.2 ± 4.1 | 15.64 ± 1.15 | 37.6 ± 221,464.0 | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| I91K | 0.31 | 3.73 ± 0.92 | 4,015.9 ± 262.1 | 1,075.53 ± 275.42 | 39.7 ± 0.3 | — | — | ||
| I91M | 0.87 | 9.04 ± 0.55 | 148.5 ± 2.8 | 16.43 ± 1.05 | 37.5 ± 0.3 | — | — | ||
| I91N | 1.50 | 9.39 ± 0.98 | 183.9 ± 5.9 | 19.59 ± 2.14 | 36.0 ± 0.2 | — | — | ||
| I91Q | 3.47 | 13.34 ± 0.58 | 209.0 ± 3.0 | 15.66 ± 0.72 | 37.8 ± 2,701,870.0 | — | — | ||
| ► | I91R | 0.53 ± 0.37 | 7.38 ± 1.79 | 239.8 ± 13.3 | 33.82 ± 5.66 | 37.6 ± 0.2 | — | -8.0 | |
| I91S | 1.40 | 10.13 ± 3.43 | 299.2 ± 29.0 | 29.54 ± 10.41 | 36.8 ± 0.2 | — | — | ||
| I91V | 1.88 | 10.44 ± 1.37 | 22.5 ± 0.9 | 2.16 ± 0.30 | 37.9 ± 88,473.6 | — | — | ||
| I91Y | 2.71 | 10.90 ± 0.60 | 248.8 ± 4.3 | 22.81 ± 1.31 | 35.8 ± 0.2 | — | — | ||
| H101R | * | 10.62 ± 0.53 | 868.1 ± 13.0 | 81.83 ± 4.33 | 40.1 ± 0.1 | 48.2 ± 0.0 | — | ||
| I106A | 1.03 | 5.15 ± 0.34 | 732.2 ± 13.0 | 142.12 ± 9.74 | — | — | — | ||
| ► | L108E | 2.77 ± 0.93 | 6.39 ± 0.81 | 387.5 ± 11.1 | 68.79 ± 5.53 | — | 43.3 ± 0.0 | -0.5 | |
| L108M | 2.35 | 4.14 ± 0.35 | 391.2 ± 8.6 | 94.54 ± 8.36 | — | 47.3 ± 0.1 | 0.6 | ||
| L108N | 2.91 | 4.27 ± 0.67 | 330.4 ± 13.4 | 77.38 ± 12.49 | — | 44.8 ± 0.0 | 0.0 | ||
| L108S | 0.26 | 2.76 ± 0.50 | 95.4 ± 4.1 | 34.55 ± 6.39 | 38.1 ± 0.5 | — | -1.1 | ||
| ► | M114S | 0.93 ± 0.25 | 9.80 ± 0.84 | 153.1 ± 4.9 | 13.53 ± 1.21 | — | 40.0 ± 34.1 | 5.0 | |
| ► | T116I | 1.99 ± 1.06 | 21.67 ± 8.57 | 310.5 ± 77.4 | 14.45 ± 10.02 | — | 43.5 ± 0.3 | 4.2 | |
| T116S | 1.61 | 2.21 ± 0.44 | 138.1 ± 6.3 | 62.58 ± 12.74 | — | 43.1 ± 0.1 | 4.0 | ||
| Y118A | 0.31 | 11.90 ± 1.18 | 11.2 ± 0.4 | 0.94 ± 0.10 | — | 46.0 ± 0.1 | — | ||
| ► | Y118F | 2.03 ± 1.69 | 0.87 ± 0.86 | 108.4 ± 20.7 | 124.90 ± 126.20 | — | 44.5 ± 0.6 | 3.2 | |
| H119A | * | 15.10 ± 3.36 | 116.9 ± 8.7 | 7.79 ± 1.73 | 40.2 ± 0.2 | — | — | ||
| ► | H119D | 0.39 ± 0.17 | — | — | 1.48 ± 0.23 | — | — | 3.9 | |
| H119E | — | — | — | — | — | — | — | ||
| H119M | 0.55 | 17.02 ± 4.07 | 9.8 ± 0.8 | 0.57 ± 0.15 | — | — | — | ||
| H119N | * | 23.22 ± 2.20 | 2.2 ± 0.0 | — | 40.3 ± 0.6 | — | — | ||
| H119Q | 0.80 | 67.26 ± 12.70 | 153.1 ± 16.0 | 2.28 ± 0.49 | — | 45.7 ± 0.6 | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | H119T | 1.09 ± 0.25 | 66.09 ± 31.10 | 20.7 ± 5.2 | 0.28 ± 0.12 | — | 45.1 ± 0.5 | 3.1 | |
| H119V | 0.43 | 44.36 ± 5.88 | 5.2 ± 0.3 | 0.12 ± 0.02 | — | — | — | ||
| W120A | — | — | — | — | — | — | — | ||
| ► | W120D | 0.21 ± 0.03 | 13.94 ± 4.28 | 93.9 ± 12.6 | 4.31 ± 2.14 | 35.6 ± 0.0 | — | 5.9 | |
| ► | W120E | 0.52 ± 0.41 | 2.02 ± 0.76 | 8.9 ± 0.6 | 2.25 ± 1.20 | 40.9 ± 0.1 | 44.8 ± 0.3 | 5.6 | |
| W120F | * | 16.08 ± 2.07 | 387.5 ± 17.3 | 24.03 ± 3.25 | 41.7 ± 0.4 | 50.7 ± 0.0 | — | ||
| W120G | 0.38 | — | — | 0.12 ± 0.00 | — | — | — | ||
| W120H | * | 89.18 ± 4.31 | 69.3 ± 2.2 | 0.87 ± 0.00 | 40.1 ± 0.1 | 46.2 ± 0.0 | — | ||
| W120K | 0.25 | 56.50 ± 10.55 | 1.7 ± 0.2 | 0.03 ± 0.01 | — | — | — | ||
| W120L | 1.43 | — | — | 0.61 ± 0.02 | 37.6 ± 633,161.0 | — | — | ||
| W120M | 1.46 | 50.86 ± 3.20 | 100.2 ± 3.2 | 1.97 ± 0.14 | — | 43.9 ± 0.1 | 5.2 | ||
| ► | W120N | 0.59 ± 0.30 | 11.72 ± 8.19 | 3.0 ± 0.4 | 2.56 ± 1.49 | — | 48.2 ± 0.1 | 4.6 | |
| W120P | — | — | — | — | — | — | — | ||
| W120Q | 0.63 | — | — | 0.29 ± 0.02 | — | — | — | ||
| ► | W120S | 0.56 ± 0.18 | 13.51 ± 2.65 | 1.2 ± 0.1 | 0.10 ± 0.01 | 40.3 ± 0.9 | — | 0.0 | |
| W120T | 0.46 | 0.37 ± 0.25 | 0.4 ± 0.0 | 1.01 ± 0.69 | — | 47.0 ± 2.1 | -0.2 | ||
| ► | W120Y | 3.68 ± 1.31 | 20.53 ± 2.92 | 4.6 ± 0.3 | 0.26 ± 0.05 | 34.8 ± 0.2 | 50.2 ± 1.2 | 11.1 | |
| D121F | — | — | — | — | — | — | — | ||
| ► | L122Q | 0.44 ± 0.24 | 2.46 ± 0.60 | 450.5 ± 15.3 | 169.36 ± 21.40 | 36.6 ± 0.2 | — | 8.0 | |
| P123D | — | — | — | — | — | — | 21.2 | ||
| Q124D | 0.75 | 4.60 ± 0.46 | 193.8 ± 4.9 | 42.15 ± 4.39 | 36.2 ± 0.4 | — | 10.1 | ||
| Q124G | 0.25 | 6.17 ± 0.27 | 71.3 ± 0.9 | 11.55 ± 0.52 | 36.6 ± 0.2 | — | — | ||
| Q124I | 1.74 | 3.43 ± 0.22 | 152.5 ± 2.3 | 44.49 ± 2.96 | 40.4 ± 0.7 | — | 5.4 | ||
| ► | Q124P | 0.98 ± 0.23 | 5.00 ± 0.37 | 263.9 ± 5.5 | 53.02 ± 4.35 | 39.5 ± 0.4 | — | 336.0 | |
| Q124S | 0.61 | 5.66 ± 0.77 | 141.2 ± 5.3 | 24.94 ± 3.52 | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | Q124T | 0.75 ± 0.54 | 5.54 ± 0.49 | 157.7 ± 3.8 | 28.40 ± 2.58 | 38.0 ± 0.6 | — | 7.5 | |
| W125F | 1.60 | 4.77 ± 0.44 | 47.6 ± 1.1 | 9.97 ± 0.96 | 36.8 ± 0.4 | — | 6.3 | ||
| W125R | 0.18 | 3.71 ± 0.99 | 62.4 ± 4.1 | 16.81 ± 4.60 | 41.3 ± 1.5 | — | 10.1 | ||
| E127D | 0.22 | 2.42 ± 0.32 | 136.6 ± 4.3 | 56.50 ± 7.77 | 39.2 ± 0.3 | — | — | ||
| ► | D128E | 0.35 ± 0.12 | 6.32 ± 0.70 | 561.0 ± 15.8 | 91.96 ± 9.26 | 39.1 ± 0.2 | — | 1.8 | |
| E129Q | 0.90 | 5.17 ± 0.46 | 348.9 ± 8.3 | 67.49 ± 6.21 | — | 43.4 ± 0.0 | -1.1 | ||
| T133Y | 3.22 | 1.58 ± 0.16 | 248.5 ± 6.2 | 157.21 ± 15.94 | — | 44.5 ± 0.2 | 6.8 | ||
| Q139E | 4.17 | 6.47 ± 1.12 | 754.0 ± 35.2 | 116.47 ± 20.92 | — | 47.7 ± 1.3 | -0.6 | ||
| ► | Q139K | 1.94 ± 1.74 | 5.04 ± 0.66 | 168.5 ± 6.1 | 44.04 ± 6.77 | — | 45.0 ± 0.2 | -2.4 | |
| K142R | 0.54 | 4.57 ± 1.26 | 60.2 ± 4.3 | 13.17 ± 3.74 | — | — | — | ||
| Y144W | 1.70 | 2.48 ± 0.47 | 280.5 ± 12.4 | 113.00 ± 22.09 | — | 43.8 ± 0.4 | -8.8 | ||
| ► | A145C | 2.42 | 4.02 ± 0.28 | 368.6 ± 6.5 | 91.74 ± 6.52 | — | 44.3 ± 0.1 | 3.0 | |
| A145E | 2.62 | 4.95 ± 0.38 | 295.1 ± 6.0 | 59.61 ± 4.74 | 40.8 ± 0.1 | — | — | ||
| M149I | 2.60 | 9.03 ± 0.70 | 377.2 ± 8.9 | 41.79 ± 3.38 | — | 42.2 ± 0.1 | 88.5 | ||
| ► | D150V | 1.33 ± 0.95 | 2.85 ± 0.61 | 128.4 ± 8.5 | 48.13 ± 12.79 | 39.6 ± 0.3 | 45.1 ± 0.2 | 12.7 | |
| E154A | 0.27 | 30.92 ± 13.46 | 112.2 ± 20.0 | 3.63 ± 1.71 | — | — | 1.6 | ||
| E154D | * | 3.46 ± 0.76 | 720.9 ± 39.0 | 208.26 ± 47.19 | 38.8 ± 0.3 | 49.5 ± 0.0 | — | ||
| E154F | 0.72 | 6.49 ± 0.56 | 2,334.7 ± 61.1 | 359.48 ± 32.42 | — | — | — | ||
| N157D | 0.72 | 6.32 ± 0.56 | 330.1 ± 8.2 | 52.25 ± 4.80 | — | 47.5 ± 0.0 | -1.0 | ||
| N157H | 2.06 | 2.17 ± 0.46 | 344.9 ± 16.7 | 159.00 ± 34.33 | — | 44.1 ± 0.0 | -0.4 | ||
| N157T | 0.26 | 2.55 ± 0.59 | 19.7 ± 1.1 | 7.73 ± 1.83 | — | — | — | ||
| N157Y | 1.15 | 7.39 ± 0.93 | 1,508.5 ± 51.8 | 204.18 ± 26.62 | 40.8 ± 0.2 | — | -1.6 | ||
| W158N | 2.79 | 4.95 ± 0.18 | 850.8 ± 7.9 | 171.78 ± 6.35 | — | — | — | ||
| N160A | 3.59 | 14.26 ± 1.61 | 402.4 ± 15.5 | 28.22 ± 3.37 | — | 44.1 ± 0.3 | 2.4 | ||
| N160C | 4.07 | 4.95 ± 0.66 | 499.7 ± 17.8 | 101.02 ± 14.02 | — | 42.9 ± 0.1 | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| N160H | 0.99 | 14.81 ± 0.91 | 144.3 ± 3.0 | 9.74 ± 0.63 | — | 41.2 ± 0.0 | 20.6 | ||
| N160S | 3.61 | 6.71 ± 0.56 | 393.5 ± 9.3 | 58.64 ± 5.07 | — | 42.5 ± 0.1 | — | ||
| ► | N160V | 1.77 ± 0.61 | 6.09 ± 1.22 | 560.4 ± 32.8 | 96.39 ± 27.65 | 38.9 ± 118,735.5 | — | 4.2 | |
| N163A | * | 11.95 ± 0.91 | 6.5 ± 0.0 | 0.65 ± 0.00 | 41.2 ± 0.3 | 44.4 ± 0.0 | — | ||
| N163C | * | 5.42 ± 0.32 | 21.6 ± 0.0 | 3.90 ± 0.22 | 38.0 ± 0.3 | 40.5 ± 0.0 | — | ||
| N163D | * | 15.19 ± 1.41 | 10.8 ± 0.0 | 0.65 ± 0.00 | 38.5 ± 0.3 | — | — | ||
| N163E | — | — | — | — | — | — | — | ||
| N163G | 2.38 | 9.68 ± 0.47 | 2.4 ± 0.0 | 0.25 ± 0.01 | — | 41.7 ± 0.0 | 6.0 | ||
| N163H | * | — | — | — | — | 43.8 ± 0.1 | 21.9 | ||
| N163K | — | — | — | — | — | — | — | ||
| N163Q | 0.10 | 7.01 ± 4.09 | 1.1 ± 0.2 | 0.16 ± 0.10 | — | — | — | ||
| N163S | 0.30 | 2.67 ± 0.41 | 1,758.0 ± 62.2 | 658.99 ± 104.13 | 38.1 ± 0.5 | — | — | ||
| ► | N163T | 0.09 ± 0.06 | 23.01 ± 2.67 | 4.8 ± 0.2 | 0.21 ± 0.03 | — | — | 4.5 | |
| E164A | * | 1.01 ± 0.17 | — | — | 46.0 ± 0.1 | 53.9 ± 0.0 | — | ||
| E164G | — | — | — | — | — | — | — | ||
| E164R | — | — | — | — | — | — | — | ||
| P165A | 0.20 | 3.73 ± 0.33 | 554.7 ± 12.5 | 148.77 ± 13.64 | 39.5 ± 0.2 | — | — | ||
| ► | Y166F | 0.32 ± 0.05 | 5.06 ± 1.13 | 7.3 ± 0.5 | 1.45 ± 0.39 | — | — | — | |
| ► | Y166H | 1.31 ± 0.03 | 18.37 ± 1.30 | 564.1 ± 14.2 | 31.86 ± 2.31 | — | 44.3 ± 0.1 | 3.4 | |
| Y166K | 0.34 | 2.24 ± 0.55 | 1.8 ± 0.1 | 0.80 ± 0.20 | — | 48.4 ± 0.5 | 11.4 | ||
| Y166P | — | — | — | — | — | — | — | ||
| Y166S | 1.64 | 6.67 ± 1.04 | 43.3 ± 1.9 | 6.49 ± 1.05 | — | — | 8.2 | ||
| Y166W | 1.90 | 7.58 ± 0.52 | 9.7 ± 0.2 | 1.28 ± 0.09 | — | — | — | ||
| C167A | * | 14.56 ± 1.27 | 391.8 ± 10.8 | 26.84 ± 2.38 | 39.8 ± 0.5 | 51.1 ± 0.0 | — | ||
| C167D | 1.60 | 57.52 ± 10.92 | 87.6 ± 8.8 | 1.52 ± 0.33 | — | 46.4 ± 0.1 | 7.6 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| C167E | 0.69 | 5.14 ± 0.20 | 183.5 ± 1.9 | 35.68 ± 1.44 | — | — | — | ||
| ► | C167H | 0.36 | 3.73 ± 0.63 | 168.2 ± 6.2 | 45.12 ± 7.79 | — | — | — | |
| ► | C167N | 1.46 ± 0.84 | 15.37 ± 1.75 | 385.9 ± 15.7 | 71.22 ± 28.33 | 40.2 ± 0.2 | 44.0 ± 0.2 | 5.4 | |
| C167Q | * | 4.92 ± 0.19 | 413.5 ± 4.3 | 84.00 ± 3.46 | 38.9 ± 0.1 | 48.2 ± 0.0 | — | ||
| ► | C167S | 3.71 ± 1.43 | 71.46 ± 9.54 | 17.0 ± 1.3 | 0.18 ± 0.03 | — | 48.0 ± 0.2 | 3.0 | |
| ► | C167T | 0.77 ± 0.57 | 2.06 ± 0.21 | 344.5 ± 8.4 | 83.71 ± 12.22 | 39.1 ± 0.6 | — | 10.9 | |
| A168N | 0.42 | 7.31 ± 0.40 | 157.0 ± 2.4 | 21.46 ± 1.23 | — | — | — | ||
| S169A | 1.35 | 4.33 ± 0.45 | 515.6 ± 14.8 | 119.07 ± 12.78 | — | 44.8 ± 0.5 | -2.2 | ||
| S169D | 0.13 | 68.87 ± 91.89 | 1.6 ± 1.1 | 0.02 ± 0.03 | — | — | — | ||
| S169N | 0.28 | 10.12 ± 5.32 | 1.6 ± 0.2 | 0.16 ± 0.09 | — | — | — | ||
| ► | I170A | 0.40 ± 0.03 | 2.90 ± 0.17 | 463.5 ± 6.8 | 152.12 ± 9.15 | 37.4 ± 0.1 | — | 4.8 | |
| I170Y | 0.86 | 4.82 ± 0.46 | 432.8 ± 11.0 | 89.75 ± 8.93 | 35.0 ± 0.2 | — | 6.7 | ||
| L171A | * | 11.09 ± 0.42 | 662.4 ± 6.5 | 59.75 ± 2.38 | 38.8 ± 0.5 | 42.9 ± 0.0 | — | ||
| ► | L171F | 5.30 ± 2.22 | 3.41 ± 0.46 | 229.3 ± 5.0 | 83.41 ± 8.37 | — | 50.0 ± 0.0 | 1.6 | |
| L171H | 3.26 | 7.23 ± 0.81 | 256.6 ± 8.3 | 35.48 ± 4.14 | — | 46.7 ± 0.1 | — | ||
| ► | L171I | 0.96 ± 0.40 | 10.55 ± 2.11 | 323.1 ± 17.3 | 35.33 ± 3.27 | — | 43.9 ± 0.1 | 39.3 | |
| L171M | 0.79 | 3.42 ± 0.21 | 247.9 ± 3.7 | 72.51 ± 4.49 | — | 45.5 ± 0.1 | — | ||
| ► | L171Q | 1.20 ± 0.07 | 5.37 ± 0.52 | 315.9 ± 12.3 | 105.54 ± 18.25 | — | 43.0 ± 0.1 | 3.9 | |
| L171R | * | 3.36 ± 0.23 | 331.2 ± 6.5 | 98.50 ± 6.93 | 38.8 ± 0.3 | 44.6 ± 0.0 | — | ||
| ► | L171Y | 1.24 ± 0.33 | 1.38 ± 0.41 | 332.7 ± 15.3 | 263.28 ± 48.14 | 37.2 ± 0.1 | 42.2 ± 0.0 | -0.4 | |
| ► | Y173L | 0.25 ± 0.22 | 2.09 ± 0.15 | 145.4 ± 5.3 | 92.27 ± 15.72 | — | — | 5.5 | |
| T175A | 0.38 | 10.60 ± 0.79 | 425.0 ± 9.7 | 40.10 ± 3.13 | — | — | — | ||
| T175C | 0.33 | 14.14 ± 1.21 | 2,190.6 ± 61.4 | 154.88 ± 13.95 | — | — | — | ||
| ► | T175H | 0.07 ± 0.03 | 2.55 ± 0.67 | 199.6 ± 13.2 | 127.56 ± 178.06 | 37.6 ± 1.0 | — | 10.0 | |
| T175K | 2.03 | 6.21 ± 0.54 | 708.5 ± 16.7 | 114.14 ± 10.27 | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| T175M | 2.50 | 5.04 ± 0.72 | 283.7 ± 12.1 | 56.30 ± 8.37 | — | 43.1 ± 0.1 | 0.3 | ||
| T175R | * | 3.59 ± 0.15 | 655.9 ± 6.5 | 182.71 ± 7.79 | 38.1 ± 0.1 | — | — | ||
| ► | E177A | 3.94 ± 2.19 | 7.16 ± 0.56 | 464.6 ± 7.9 | 72.08 ± 4.48 | 37.5 ± 0.2 | 47.0 ± 0.4 | 3.1 | |
| E177C | 1.52 | 6.13 ± 0.26 | 70.8 ± 0.8 | 11.55 ± 0.51 | — | — | — | ||
| ► | E177D | 1.95 ± 0.02 | 5.04 ± 0.58 | 404.1 ± 12.5 | 80.17 ± 9.60 | 37.9 ± 78,411.8 | — | — | |
| E177I | — | — | — | — | — | — | — | ||
| E177K | * | 6.19 ± 0.30 | 454.6 ± 6.5 | 73.39 ± 3.68 | 36.7 ± 0.3 | — | — | ||
| E177L | * | 7.48 ± 0.36 | 549.9 ± 8.7 | 73.60 ± 3.68 | 39.6 ± 0.4 | — | — | ||
| E177Q | 0.58 | 8.17 ± 0.46 | 799.4 ± 13.0 | 97.81 ± 5.78 | — | — | — | ||
| ► | E177T | 2.41 ± 0.31 | 7.59 ± 0.87 | 382.0 ± 11.1 | 49.04 ± 5.71 | — | 43.6 ± 0.1 | — | |
| ► | E177W | 0.29 ± 0.09 | 10.32 ± 1.33 | 61.0 ± 1.5 | 9.64 ± 0.92 | 39.6 ± 0.3 | — | 5.2 | |
| H178A | * | 7.67 ± 0.73 | 93.1 ± 2.2 | 12.12 ± 1.08 | 39.6 ± 0.4 | — | — | ||
| ► | H178C | 0.10 ± 0.06 | 3.90 ± 0.31 | 93.3 ± 4.0 | 45.53 ± 17.40 | — | — | 3.4 | |
| ► | H178F | 0.28 ± 0.16 | 4.72 ± 0.34 | 171.2 ± 4.6 | 46.35 ± 5.58 | — | 40.5 ± 0.1 | -3.4 | |
| H178K | 0.67 | 25.15 ± 1.69 | 22.5 ± 0.6 | 0.90 ± 0.06 | — | — | 8.9 | ||
| ► | H178N | 1.16 ± 1.06 | 4.80 ± 0.68 | 340.4 ± 14.2 | 107.79 ± 42.16 | 39.4 ± 0.2 | — | 3.3 | |
| H178Q | 0.17 | 8.73 ± 0.50 | 231.4 ± 4.0 | 26.51 ± 1.59 | — | — | 2.6 | ||
| ► | H178R | 0.09 | 4.96 ± 0.48 | 48.9 ± 1.4 | 9.85 ± 1.00 | — | — | — | |
| H178S | 0.09 | 14.86 ± 0.61 | 51.0 ± 0.7 | 3.43 ± 0.15 | — | — | — | ||
| ► | H178W | 1.02 ± 0.30 | 7.00 ± 1.17 | 331.7 ± 18.5 | 89.44 ± 20.44 | — | — | 160.8 | |
| H178Y | 0.16 | 4.73 ± 0.24 | 4.0 ± 0.1 | 0.84 ± 0.04 | — | — | -1.2 | ||
| H182K | 5.12 | 4.43 ± 0.35 | 377.2 ± 7.7 | 85.12 ± 6.87 | — | 50.0 ± 0.0 | -0.8 | ||
| H182Q | 5.63 | 5.81 ± 0.38 | 412.3 ± 8.1 | 71.02 ± 4.90 | — | 49.5 ± 0.7 | 2.0 | ||
| H182R | 0.68 | 10.04 ± 0.98 | 720.0 ± 21.1 | 71.68 ± 7.28 | — | — | — | ||
| H182S | 1.18 | 4.26 ± 0.23 | 333.9 ± 8.3 | 78.41 ± 4.66 | — | 48.5 ± 0.7 | 2.7 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| H182T | 4.02 | 3.72 ± 0.29 | 191.0 ± 9.4 | 51.36 ± 4.74 | — | 49.3 ± 1.2 | 8.1 | ||
| H182V | — | — | — | — | — | — | 6.6 | ||
| H182Y | 2.90 | 8.33 ± 2.15 | 528.0 ± 53.4 | 63.39 ± 17.61 | — | 47.2 ± 0.3 | 3.0 | ||
| E183D | 0.00 | 1.09 ± 0.12 | 885.3 ± 21.5 | 813.55 ± 90.21 | 39.6 ± 0.5 | — | 1.8 | ||
| E183H | 0.86 | 6.40 ± 0.55 | 355.9 ± 8.3 | 55.64 ± 4.93 | — | — | — | ||
| E183T | 4.10 | 4.50 ± 0.25 | 581.3 ± 8.6 | 129.17 ± 7.51 | — | — | — | ||
| N184D | 1.31 | 3.84 ± 0.44 | 1,109.4 ± 32.1 | 288.79 ± 33.88 | 40.4 ± 0.3 | — | -0.2 | ||
| E187A | 8.07 | 5.94 ± 0.37 | 384.4 ± 8.6 | 64.76 ± 4.25 | — | 51.2 ± 0.6 | 2.3 | ||
| E187F | 6.13 | 5.32 ± 0.22 | 40.9 ± 0.5 | 7.69 ± 0.34 | — | 50.8 ± 0.6 | 1.8 | ||
| E187T | 2.84 | 6.15 ± 1.07 | 404.9 ± 20.1 | 65.81 ± 11.85 | — | 49.2 ± 0.8 | 5.4 | ||
| A192S | * | 5.09 ± 0.18 | 775.0 ± 8.7 | 152.19 ± 5.63 | 39.2 ± 0.3 | — | — | ||
| H194G | 0.20 | 1.59 ± 0.30 | 91.5 ± 4.4 | 57.56 ± 11.06 | — | — | 7.0 | ||
| ► | H194L | 0.24 ± 0.05 | 2.77 ± 0.38 | 133.3 ± 5.3 | 58.22 ± 11.21 | 37.4 ± 0.2 | — | 28.9 | |
| ► | H194N | 1.07 ± 0.22 | 4.51 ± 0.43 | 168.8 ± 4.2 | 37.42 ± 3.70 | 38.9 ± 0.4 | — | — | |
| H194Q | 0.16 | 6.08 ± 0.86 | 78.8 ± 3.1 | 12.96 ± 1.90 | 37.2 ± 0.3 | — | — | ||
| H194V | 1.04 | 4.27 ± 0.53 | 194.4 ± 6.3 | 45.52 ± 5.88 | 39.9 ± 0.5 | — | 22.1 | ||
| ► | H194Y | 0.75 ± 0.01 | 5.62 ± 0.80 | 186.8 ± 6.7 | 33.86 ± 4.11 | 38.9 ± 0.3 | — | — | |
| C198A | 3.03 | 4.73 ± 0.47 | 82.9 ± 2.2 | 17.52 ± 1.79 | 39.3 ± 0.2 | — | -0.9 | ||
| C198D | — | — | — | — | — | — | 6.8 | ||
| C198F | — | — | — | — | — | — | 287.9 | ||
| C198G | 0.93 | 1.59 ± 0.34 | 18.0 ± 0.8 | 11.33 ± 2.49 | 37.2 ± 0.4 | — | 4.1 | ||
| C198K | 1.76 | 4.96 ± 0.18 | 84.9 ± 0.8 | 17.13 ± 0.65 | 36.7 ± 1.3 | — | 55.6 | ||
| C198L | 0.68 | — | — | — | — | — | 33.2 | ||
| C198M | 1.24 | 4.93 ± 0.67 | 80.9 ± 2.9 | 16.42 ± 2.31 | 36.1 ± 0.4 | — | 42.9 | ||
| C198N | — | — | — | — | — | — | 12.0 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| C198Q | 0.65 | 1.57 ± 0.32 | 2.8 ± 0.1 | 1.78 ± 0.38 | 40.9 ± 1.1 | — | 16.7 | ||
| C198S | 1.98 | 2.75 ± 0.39 | 113.1 ± 3.8 | 41.19 ± 5.96 | 38.6 ± 0.3 | — | 2.0 | ||
| C198T | 5.49 | 2.27 ± 0.24 | 132.1 ± 3.2 | 58.08 ± 6.26 | 35.9 ± 0.3 | — | 3.2 | ||
| S203V | 0.98 | 3.05 ± 0.46 | 1,281.5 ± 47.5 | 419.79 ± 65.64 | 44.3 ± 0.3 | — | -4.1 | ||
| ► | T212E | 1.21 ± 0.07 | 4.93 ± 0.62 | 384.1 ± 13.2 | 77.91 ± 10.34 | — | 46.5 ± 0.2 | -0.2 | |
| I217S | 1.08 | 5.46 ± 0.66 | 1,280.4 ± 42.3 | 234.72 ± 29.55 | 42.4 ± 0.1 | — | 6.4 | ||
| T218A | * | 6.51 ± 0.94 | 381.0 ± 15.2 | 58.45 ± 8.66 | 40.1 ± 0.1 | 47.5 ± 0.0 | — | ||
| T218G | 1.78 | 6.66 ± 1.34 | 142.0 ± 8.0 | 21.33 ± 4.45 | — | 43.0 ± 0.5 | 8.3 | ||
| ► | T218Q | 2.72 ± 1.68 | 5.49 ± 0.46 | 376.9 ± 8.6 | 34.33 ± 4.21 | — | 46.3 ± 0.7 | 22.8 | |
| ► | T218S | 2.06 ± 0.95 | 22.52 ± 2.93 | 352.7 ± 18.2 | 18.48 ± 3.21 | 41.0 ± 0.2 | 47.5 ± 0.3 | — | |
| L219M | 0.82 | 5.58 ± 0.63 | 248.5 ± 7.6 | 44.54 ± 5.18 | — | 44.5 ± 0.3 | — | ||
| ► | L219Q | 1.02 ± 0.52 | 6.36 ± 2.47 | 719.5 ± 21.5 | 104.07 ± 14.53 | — | — | 6.6 | |
| L219R | 3.02 | 2.79 ± 0.26 | 278.9 ± 6.2 | 100.01 ± 9.44 | 36.6 ± 0.3 | — | — | ||
| L219V | 1.38 | 3.46 ± 0.19 | 225.4 ± 3.1 | 65.05 ± 3.75 | 37.2 ± 0.1 | — | — | ||
| ► | N220A | 3.07 ± 3.46 | 15.35 ± 8.87 | 154.8 ± 9.1 | 13.59 ± 1.46 | 38.8 ± 0.2 | 43.6 ± 0.3 | 1.7 | |
| ► | N220C | 1.07 ± 0.25 | 4.15 ± 0.51 | 224.1 ± 13.1 | 90.44 ± 33.48 | 36.6 ± 0.3 | — | — | |
| ► | N220D | 1.35 ± 0.19 | 3.21 ± 0.34 | 54.6 ± 3.1 | 34.75 ± 9.98 | 35.3 ± 0.2 | — | — | |
| ► | N220E | 2.36 ± 2.02 | 51.99 ± 9.51 | 202.2 ± 26.7 | 3.06 ± 0.73 | 36.3 ± 0.2 | — | 5.6 | |
| ► | N220F | 1.85 ± 0.73 | 1.17 ± 0.20 | 397.9 ± 9.8 | 546.89 ± 60.45 | 37.9 ± 14,182.7 | — | -2.0 | |
| ► | N220G | 10.31 | 31.79 ± 2.80 | 92.5 ± 3.1 | 2.83 ± 0.31 | 38.9 ± 0.3 | 47.4 ± 0.0 | 5.2 | |
| ► | N220H | 7.46 | 4.81 ± 0.23 | 58.8 ± 0.2 | 11.61 ± 0.63 | 39.1 ± 0.3 | 46.8 ± 0.0 | 0.0 | |
| N220I | * | — | — | — | 36.7 ± 0.1 | — | — | ||
| ► | N220K | 3.11 ± 0.45 | 2.21 ± 0.41 | 351.6 ± 15.0 | 159.19 ± 30.49 | 40.0 ± 0.4 | — | — | |
| ► | N220L | 3.30 ± 1.72 | 10.51 ± 2.96 | 368.6 ± 20.7 | 65.72 ± 10.00 | 37.2 ± 1.0 | — | — | |
| ► | N220M | 4.02 ± 3.49 | 1.52 ± 0.12 | 226.6 ± 5.1 | 144.15 ± 14.76 | 36.3 ± 0.2 | — | 1.0 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| N220P | 3.79 | 9.45 ± 0.46 | 77.1 ± 1.1 | 8.15 ± 0.41 | 35.6 ± 0.2 | — | 53.3 | ||
| ► | N220Q | 3.34 ± 3.05 | 5.28 ± 0.97 | 83.1 ± 6.5 | 17.36 ± 5.35 | 36.4 ± 0.2 | — | 2.6 | |
| ► | N220R | 3.91 ± 3.32 | 10.03 ± 1.41 | 145.1 ± 4.4 | 22.54 ± 2.61 | 40.3 ± 0.4 | 46.9 ± 0.0 | -9.4 | |
| ► | N220S | 1.18 ± 0.50 | 32.82 ± 2.17 | 68.9 ± 1.8 | 3.27 ± 0.14 | 40.4 ± 0.2 | 47.2 ± 0.1 | — | |
| ► | N220T | 4.29 | 11.90 ± 1.23 | 0.4 ± 0.0 | 0.04 ± 0.00 | 39.7 ± 0.2 | — | — | |
| ► | N220V | 1.30 | 2.98 ± 0.42 | 112.1 ± 3.8 | 37.68 ± 5.46 | 36.6 ± 0.3 | — | 54.0 | |
| ► | N220W | 0.69 ± 0.11 | 0.84 ± 0.22 | 165.3 ± 11.3 | 169.64 ± 46.01 | — | — | — | |
| ► | N220Y | 7.90 ± 4.67 | 2.21 ± 0.12 | 349.7 ± 3.6 | 544.03 ± 42.59 | 40.8 ± 0.1 | 49.2 ± 0.0 | -2.3 | |
| M221A | * | 6.25 ± 0.60 | 448.1 ± 13.0 | 71.66 ± 7.14 | 40.4 ± 0.3 | 47.2 ± 0.0 | — | ||
| ► | M221K | 0.20 ± 0.03 | 9.48 ± 0.59 | 114.2 ± 2.2 | 12.30 ± 0.88 | — | 46.5 ± 0.1 | 5.5 | |
| M221N | 1.50 | 5.59 ± 0.33 | 246.3 ± 3.9 | 44.07 ± 2.67 | 36.6 ± 0.1 | — | — | ||
| M221Q | 2.84 | 8.03 ± 0.20 | 1,008.5 ± 7.3 | 125.56 ± 3.32 | — | — | — | ||
| M221R | 0.24 | 14.98 ± 0.66 | 25.1 ± 0.4 | 1.67 ± 0.08 | — | — | 3.2 | ||
| E222A | * | 0.63 ± 0.15 | 73.6 ± 4.3 | 116.90 ± 28.58 | 36.9 ± 0.1 | 36.7 ± 0.0 | — | ||
| E222H | * | 8.54 ± 0.53 | 132.1 ± 2.2 | 15.37 ± 1.08 | 34.9 ± 0.2 | 40.2 ± 0.0 | — | ||
| E222K | * | 7.22 ± 0.75 | 88.8 ± 2.2 | 12.34 ± 1.30 | 39.0 ± 0.4 | — | — | ||
| E222Q | * | 12.16 ± 0.65 | 547.7 ± 8.7 | 45.03 ± 2.60 | 39.8 ± 0.3 | — | — | ||
| E222R | * | 2.48 ± 0.44 | 34.6 ± 2.2 | 14.07 ± 2.60 | 39.1 ± 0.2 | — | — | ||
| E222Y | * | 18.43 ± 3.14 | 10.8 ± 0.0 | 0.65 ± 0.22 | 37.1 ± 0.5 | — | — | ||
| ► | H223A | 1.79 ± 1.61 | 5.83 ± 1.22 | 363.0 ± 35.3 | 64.41 ± 17.19 | 37.3 ± 1.2 | 42.0 ± 0.2 | -2.5 | |
| H223S | 4.60 | 6.49 ± 0.92 | 248.1 ± 9.9 | 38.21 ± 5.61 | — | 40.0 ± 0.1 | 1.2 | ||
| P231K | 2.98 | 4.57 ± 0.42 | 349.8 ± 8.5 | 76.53 ± 7.33 | — | 44.8 ± 0.2 | 0.2 | ||
| D233I | 0.58 | 3.94 ± 0.38 | 33.7 ± 1.1 | 8.55 ± 0.88 | — | — | 16.2 | ||
| A236C | 2.40 | 3.89 ± 0.44 | 202.5 ± 5.9 | 52.09 ± 6.13 | — | 39.4 ± 0.9 | 40.3 | ||
| ► | A236D | — | — | — | — | — | — | 43.0 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | A236E | * | — | — | — | — | 51.8 ± 0.4 | 160.3 | |
| A236S | 0.13 | — | — | 0.01 ± 0.00 | — | 45.4 ± 0.8 | 6.7 | ||
| ► | A236V | 1.03 ± 0.52 | 0.45 ± 0.31 | 1.2 ± 0.2 | 2.51 ± 1.90 | 36.2 ± 1.3 | — | — | |
| R239C | 1.55 | 6.52 ± 0.41 | 161.3 ± 2.8 | 24.76 ± 1.61 | — | — | 6.7 | ||
| ► | R240A | 2.75 ± 0.95 | 12.63 ± 2.26 | 427.2 ± 18.5 | 34.48 ± 4.60 | 38.5 ± 0.2 | 44.8 ± 0.2 | 2.4 | |
| R240D | * | 10.82 ± 0.47 | 231.6 ± 4.3 | 21.43 ± 1.08 | 37.2 ± 0.0 | 45.7 ± 0.0 | — | ||
| R240E | — | — | — | — | — | — | — | ||
| ► | R240G | 0.25 ± 0.05 | 23.66 ± 2.61 | 456.7 ± 19.1 | 19.31 ± 2.28 | — | — | — | |
| ► | R240K | 2.20 | 12.92 ± 2.43 | 726.3 ± 37.5 | 64.63 ± 9.16 | 39.5 ± 0.2 | 44.0 ± 0.0 | 2.2 | |
| D241H | 0.28 | 9.97 ± 0.78 | 100.2 ± 2.4 | 10.05 ± 0.82 | — | — | — | ||
| D241R | 0.13 | 2.42 ± 0.46 | 5.1 ± 0.2 | 2.10 ± 0.41 | — | — | 1.3 | ||
| ► | F243H | 1.94 ± 1.39 | 9.59 ± 3.28 | 477.8 ± 40.3 | 148.04 ± 53.52 | 41.5 ± 0.3 | 45.9 ± 0.0 | 2.2 | |
| I244A | 0.20 | 8.50 ± 1.59 | 451.3 ± 16.6 | 53.12 ± 10.12 | 40.4 ± 0.2 | — | 5.3 | ||
| I244E | * | 5.97 ± 1.04 | 407.0 ± 19.5 | 68.19 ± 12.34 | 37.8 ± 0.1 | 45.8 ± 0.0 | — | ||
| I244K | 3.48 | 14.07 ± 0.87 | 134.4 ± 2.8 | 9.55 ± 0.63 | — | — | 4.3 | ||
| ► | I244M | 2.97 | 2.72 ± 0.59 | 236.0 ± 15.5 | 170.89 ± 111.12 | — | — | 6.8 | |
| I244N | * | 2.15 ± 0.13 | 223.0 ± 4.3 | 103.69 ± 6.49 | 36.8 ± 0.6 | 45.0 ± 0.0 | — | ||
| I244Q | 2.18 | 14.73 ± 0.94 | 1,166.6 ± 24.6 | 79.20 ± 5.31 | — | — | — | ||
| I244T | 3.11 | 4.22 ± 0.17 | 681.8 ± 6.9 | 161.68 ± 6.69 | — | — | — | ||
| I244V | 1.54 | 2.04 ± 0.16 | 518.2 ± 10.0 | 254.43 ± 21.17 | — | — | — | ||
| ► | R246K | 0.77 ± 0.81 | 2.15 ± 0.33 | 467.9 ± 28.2 | 646.86 ± 284.71 | 36.8 ± 0.3 | 42.0 ± 0.4 | -0.6 | |
| W247V | 0.16 | 7.62 ± 2.81 | 4.2 ± 0.4 | 0.55 ± 0.21 | — | — | — | ||
| F248N | 0.30 | 8.99 ± 2.33 | 8.1 ± 0.6 | 0.91 ± 0.25 | — | 50.3 ± 0.7 | 6.2 | ||
| A249E | — | — | — | — | — | — | — | ||
| ► | E250D | 0.89 ± 0.12 | 2.62 ± 0.38 | 816.4 ± 29.3 | 414.04 ± 89.94 | 40.0 ± 0.5 | — | -1.2 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| F253H | 0.28 | 64.81 ± 42.05 | 297.9 ± 110.9 | 4.60 ± 3.44 | — | — | 3.6 | ||
| ► | F253I | 2.56 ± 1.15 | 12.23 ± 1.70 | 315.8 ± 9.7 | 48.72 ± 5.86 | — | 45.3 ± 0.2 | 5.9 | |
| ► | F253M | 2.17 ± 1.17 | 5.99 ± 0.39 | 448.5 ± 8.2 | 74.93 ± 5.10 | — | 44.7 ± 0.1 | 6.8 | |
| ► | F253Y | 0.69 ± 0.46 | 6.86 ± 0.56 | 275.6 ± 6.6 | 40.53 ± 3.70 | — | 40.9 ± 0.1 | 1.4 | |
| D260E | 0.88 | 10.56 ± 0.84 | 1,349.4 ± 33.7 | 127.83 ± 10.64 | — | — | — | ||
| M261D | — | — | — | — | — | — | — | ||
| W264H | 0.27 | 10.37 ± 1.48 | 124.0 ± 6.0 | 11.96 ± 1.80 | — | 45.2 ± 0.0 | 0.0 | ||
| W264Y | 0.16 | 4.66 ± 2.54 | 24.4 ± 3.5 | 5.23 ± 2.96 | — | — | — | ||
| Y265F | 0.30 | 11.19 ± 3.84 | 5.5 ± 0.6 | 0.49 ± 0.18 | 34.5 ± 0.8 | 30.7 ± 18.7 | -0.6 | ||
| T267E | 2.77 | 4.58 ± 0.47 | 422.5 ± 11.4 | 92.19 ± 9.79 | — | 47.6 ± 0.2 | -1.3 | ||
| Y268K | 1.19 | 3.03 ± 0.36 | 1,237.6 ± 35.5 | 408.00 ± 49.49 | 39.7 ± 0.1 | — | 9.2 | ||
| L269I | 0.21 | 3.77 ± 0.53 | 176.9 ± 6.2 | 46.90 ± 6.80 | 40.4 ± 0.6 | — | 0.4 | ||
| ► | L269V | 0.53 ± 0.10 | 4.17 ± 0.73 | 898.7 ± 40.4 | 215.66 ± 38.85 | 40.0 ± 1.0 | — | 21.6 | |
| ► | N270G | 1.14 ± 0.16 | 7.15 ± 1.88 | 1,307.1 ± 97.1 | 246.14 ± 33.92 | 40.6 ± 0.2 | — | 3.2 | |
| ► | N270W | 1.11 ± 1.00 | 3.30 ± 0.47 | 345.6 ± 13.9 | 177.72 ± 52.16 | 39.7 ± 0.4 | 42.2 ± 0.1 | 8.6 | |
| ► | G271D | 0.46 ± 0.15 | 1.41 ± 0.22 | 592.0 ± 21.7 | 376.28 ± 63.75 | 37.4 ± 0.2 | — | 4.7 | |
| L272F | 1.23 | 0.80 ± 0.78 | 2.4 ± 0.5 | 3.01 ± 2.99 | — | — | 80.9 | ||
| F274N | 0.70 | 1.96 ± 0.22 | 156.5 ± 4.0 | 79.66 ± 9.10 | 34.9 ± 0.1 | — | — | ||
| Q276K | 0.74 | 3.48 ± 0.47 | 424.1 ± 14.2 | 121.94 ± 16.83 | 34.2 ± 0.9 | — | 4.6 | ||
| G278V | 3.54 | 5.16 ± 1.58 | 463.0 ± 36.2 | 89.76 ± 28.41 | — | — | — | ||
| M280I | 0.54 | 5.70 ± 0.43 | 451.7 ± 9.1 | 79.19 ± 6.22 | — | — | — | ||
| ► | D288E | 2.35 ± 0.06 | 4.26 ± 0.43 | 520.3 ± 13.8 | 114.33 ± 11.87 | — | 44.0 ± 0.1 | 3.6 | |
| F289A | — | — | — | — | — | — | 7.0 | ||
| L290V | 0.39 | 3.65 ± 0.41 | 1,461.3 ± 41.3 | 400.17 ± 46.29 | — | 44.3 ± 0.2 | — | ||
| ► | N293A | 0.79 | 11.73 ± 1.02 | 9.3 ± 0.2 | 0.83 ± 0.04 | 40.8 ± 0.2 | 46.5 ± 0.1 | -0.4 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| N293C | * | 19.80 ± 2.08 | 8.7 ± 0.0 | 0.43 ± 0.00 | 38.9 ± 0.4 | 47.0 ± 0.0 | — | ||
| N293D | — | — | — | — | — | — | — | ||
| N293E | * | — | — | — | — | — | 18.9 | ||
| N293F | — | — | — | — | — | — | 86.3 | ||
| ► | N293G | 3.88 ± 2.24 | 24.01 ± 2.80 | 1.1 ± 0.0 | 0.04 ± 0.01 | — | — | — | |
| N293H | * | — | — | — | — | 42.1 ± 0.0 | 162.0 | ||
| N293I | 0.64 | 1.43 ± 0.28 | 1.3 ± 0.1 | 0.92 ± 0.19 | 39.6 ± 1.3 | — | 77.0 | ||
| N293K | — | — | — | — | — | — | — | ||
| N293L | — | — | — | — | — | — | 67.3 | ||
| N293M | * | — | — | — | — | — | 41.9 | ||
| N293P | — | — | — | — | — | — | — | ||
| N293Q | * | — | — | — | — | — | — | ||
| N293R | 0.37 | 0.70 ± 0.15 | 7.9 ± 0.3 | 11.25 ± 2.44 | — | — | — | ||
| ► | N293S | 4.54 ± 1.63 | 2.11 ± 0.46 | 2.8 ± 0.2 | 2.90 ± 1.39 | 41.3 ± 0.8 | 47.8 ± 0.5 | 0.6 | |
| N293T | 2.11 | 13.62 ± 1.65 | 29.2 ± 1.2 | 2.15 ± 0.27 | 38.3 ± 0.4 | 38.3 ± 0.4 | 11.8 | ||
| ► | N293V | 1.71 ± 0.35 | 5.26 ± 0.89 | 2.7 ± 0.4 | 1.23 ± 0.84 | 37.9 ± 67,349.9 | 43.4 ± 0.4 | 7.7 | |
| ► | N293W | — | — | — | — | — | — | 119.6 | |
| ► | N293Y | 0.61 | 48.85 ± 36.83 | 0.6 ± 0.2 | 0.01 ± 0.01 | — | 49.0 ± 0.8 | 87.2 | |
| Y294A | * | 4.98 ± 0.17 | 136.4 ± 2.2 | 27.49 ± 1.08 | 40.4 ± 0.2 | 43.3 ± 0.0 | — | ||
| Y294F | * | 5.99 ± 0.32 | 601.8 ± 8.7 | 100.45 ± 5.63 | 40.8 ± 0.1 | 44.0 ± 0.0 | — | ||
| Y294H | * | — | — | — | — | — | 3.2 | ||
| Y294L | — | — | — | — | — | — | — | ||
| Y295A | * | — | — | — | — | — | — | ||
| Y295G | * | — | — | — | — | — | — | ||
| Y295N | 1.59 | 18.93 ± 3.85 | 140.3 ± 10.5 | 7.41 ± 1.61 | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | T296A | 2.99 | 47.09 ± 4.63 | 0.6 ± 0.0 | 0.01 ± 0.00 | — | — | — | |
| ► | T296C | 0.60 ± 0.43 | 10.65 ± 1.48 | 248.3 ± 15.2 | 21.43 ± 4.20 | — | — | — | |
| T296E | — | — | — | — | — | — | — | ||
| T296F | 5.25 | 0.84 ± 0.14 | 226.7 ± 7.7 | 269.30 ± 46.46 | — | — | 0.1 | ||
| T296H | 0.46 | 8.23 ± 1.10 | 101.5 ± 3.9 | 12.33 ± 1.72 | — | — | — | ||
| T296R | 0.91 | 1.27 ± 0.15 | 780.7 ± 23.1 | 615.34 ± 75.45 | 42.4 ± 0.1 | — | 0.8 | ||
| T296S | 2.46 | 3.91 ± 0.83 | 297.1 ± 18.2 | 76.00 ± 16.89 | — | 43.8 ± 0.3 | 0.4 | ||
| T296W | 2.62 | 0.66 ± 0.10 | 3.0 ± 0.1 | 4.49 ± 0.73 | — | — | 3.0 | ||
| T296Y | 4.35 | 5.95 ± 0.52 | 497.2 ± 12.0 | 83.60 ± 7.60 | — | 45.4 ± 0.0 | -1.1 | ||
| R297A | 0.62 | 2.36 ± 0.49 | 52.2 ± 2.6 | 22.07 ± 4.74 | 41.8 ± 0.1 | — | 5.9 | ||
| S298E | * | 5.28 ± 0.05 | 662.4 ± 2.2 | 125.56 ± 1.30 | 39.6 ± 0.2 | — | — | ||
| S298R | 0.76 | 2.56 ± 1.14 | 435.2 ± 45.8 | 170.03 ± 77.66 | 39.4 ± 0.2 | — | — | ||
| S298T | 0.85 | 3.34 ± 0.37 | 766.1 ± 21.2 | 229.47 ± 26.45 | 39.4 ± 0.3 | — | — | ||
| ► | I299R | 4.56 | 3.48 ± 0.55 | 263.8 ± 10.5 | 75.84 ± 12.43 | — | — | 3.2 | |
| I300N | * | 4.48 ± 0.32 | 569.3 ± 10.8 | 127.07 ± 9.31 | 38.6 ± 0.5 | — | — | ||
| ► | S302A | 2.13 ± 0.03 | 4.83 ± 0.24 | 329.6 ± 4.4 | 68.28 ± 3.52 | — | 44.7 ± 0.0 | -0.8 | |
| S302D | 6.77 | 14.89 ± 1.41 | 74.6 ± 2.4 | 5.01 ± 0.50 | 39.2 ± 0.2 | — | 5.9 | ||
| ► | S302F | 7.84 ± 1.20 | 3.79 ± 0.29 | 124.9 ± 2.6 | 35.00 ± 3.80 | 39.3 ± 0.1 | — | 6.2 | |
| ► | S302H | 3.21 ± 2.02 | 4.81 ± 0.37 | 568.3 ± 14.0 | 133.50 ± 16.70 | — | — | — | |
| S302K | 6.91 | 5.72 ± 0.53 | 197.8 ± 5.0 | 34.56 ± 3.32 | 37.6 ± 0.5 | — | 9.8 | ||
| S302N | 3.27 | 5.43 ± 0.16 | 718.7 ± 5.4 | 132.24 ± 3.92 | — | — | — | ||
| ► | S302W | 5.62 ± 0.69 | 5.41 ± 0.87 | 193.1 ± 12.1 | 38.55 ± 10.43 | 38.8 ± 0.3 | — | 6.2 | |
| S302Y | 11.07 | 5.53 ± 0.48 | 153.3 ± 3.7 | 27.74 ± 2.52 | 39.9 ± 0.1 | — | 5.8 | ||
| T303K | 1.17 | 3.45 ± 0.37 | 865.0 ± 23.3 | 250.97 ± 27.94 | 42.0 ± 0.1 | — | 76.0 | ||
| A306K | 1.55 | 4.71 ± 0.54 | 698.6 ± 21.3 | 148.21 ± 17.68 | 40.7 ± 0.2 | — | -1.5 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | A306N | 1.68 ± 0.09 | 4.97 ± 0.19 | 340.9 ± 3.5 | 68.54 ± 2.68 | — | 44.6 ± 0.1 | -2.1 | |
| ► | A306Q | 2.08 | 3.75 ± 0.63 | 587.8 ± 25.0 | 156.87 ± 27.23 | 38.3 ± 0.8 | — | — | |
| ► | A306S | 1.21 ± 0.66 | 6.93 ± 0.77 | 486.2 ± 16.0 | 132.59 ± 31.90 | 40.4 ± 0.1 | 44.6 ± 0.2 | -1.6 | |
| V311D | 3.05 | 6.84 ± 0.65 | 295.0 ± 8.0 | 43.14 ± 4.25 | — | 41.8 ± 0.1 | 2.6 | ||
| ► | E312V | 0.87 ± 0.12 | 3.37 ± 0.18 | 958.4 ± 15.2 | 287.10 ± 19.44 | 38.6 ± 0.2 | — | 3.7 | |
| Q313A | 1.25 | 4.45 ± 0.57 | 603.3 ± 19.7 | 135.49 ± 17.81 | 39.6 ± 0.1 | — | — | ||
| ► | Q313C | 3.14 ± 1.54 | 5.54 ± 0.46 | 285.0 ± 5.3 | 49.25 ± 3.80 | — | — | 0.9 | |
| Q313E | 4.20 | 6.76 ± 0.86 | 413.8 ± 14.9 | 61.19 ± 8.05 | 39.7 ± 0.1 | — | — | ||
| Q313L | 3.90 | 3.58 ± 0.23 | 363.5 ± 5.8 | 101.67 ± 6.76 | 40.1 ± 0.1 | — | — | ||
| Q313N | 0.37 | 6.14 ± 0.89 | 291.1 ± 11.4 | 47.42 ± 7.10 | — | — | — | ||
| Q313P | 2.63 | 3.48 ± 0.55 | 126.7 ± 4.7 | 36.42 ± 5.92 | — | — | 4.3 | ||
| Q313R | * | 3.58 ± 0.51 | 565.0 ± 19.5 | 157.82 ± 23.16 | 42.1 ± 0.3 | — | — | ||
| Q313V | 2.53 | 8.77 ± 0.68 | 900.2 ± 20.5 | 102.63 ± 8.35 | — | — | — | ||
| V314A | 2.85 | 2.64 ± 0.29 | 182.3 ± 4.8 | 69.19 ± 7.93 | 37.9 ± 0.4 | — | 2.8 | ||
| V314D | 8.32 | 5.14 ± 0.61 | 126.2 ± 4.1 | 24.58 ± 3.05 | 38.1 ± 0.2 | — | 6.7 | ||
| V314E | 6.38 | 2.42 ± 0.34 | 125.9 ± 4.4 | 52.01 ± 7.45 | 38.0 ± 0.1 | — | 2.8 | ||
| V314F | 5.00 | 4.47 ± 0.33 | 210.1 ± 4.0 | 46.98 ± 3.54 | 37.8 ± 0.2 | — | 8.2 | ||
| V314H | 6.95 | 3.43 ± 0.33 | 227.5 ± 5.8 | 66.32 ± 6.57 | 38.2 ± 0.1 | — | 5.6 | ||
| V314K | 2.94 | 5.55 ± 0.83 | 269.1 ± 12.3 | 48.45 ± 7.60 | 36.0 ± 0.4 | — | 2.4 | ||
| ► | V314L | 8.02 ± 0.87 | 1.82 ± 0.33 | 151.2 ± 5.2 | 108.02 ± 17.69 | 38.8 ± 0.3 | — | 2.3 | |
| V314M | 7.63 | 6.76 ± 0.43 | 181.4 ± 3.3 | 26.84 ± 1.79 | 38.5 ± 0.2 | — | 4.3 | ||
| V314Q | 6.59 | 3.16 ± 0.49 | 165.6 ± 6.3 | 52.36 ± 8.31 | 38.5 ± 0.2 | — | 2.4 | ||
| V314T | 2.24 | 3.44 ± 0.37 | 164.7 ± 4.5 | 47.91 ± 5.38 | 38.5 ± 0.3 | — | 3.3 | ||
| V314W | 6.07 | 4.76 ± 0.20 | 168.1 ± 1.9 | 35.36 ± 1.58 | 37.1 ± 0.2 | — | 7.7 | ||
| ► | H315C | 0.37 | 2.98 ± 0.69 | 0.7 ± 0.0 | 0.23 ± 0.06 | 38.9 ± 0.7 | — | — | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| H315D | 0.32 | 0.68 ± 0.24 | 897.0 ± 54.3 | 1,312.84 ± 460.56 | 40.1 ± 0.1 | — | — | ||
| ► | H315E | 1.78 | 2.84 ± 0.39 | 183.7 ± 6.8 | 64.70 ± 9.19 | — | 47.2 ± 0.1 | -1.2 | |
| H315I | 3.82 | 6.73 ± 0.32 | 588.1 ± 7.6 | 87.37 ± 4.25 | — | — | — | ||
| H315K | 0.38 | 6.14 ± 0.56 | 36.6 ± 0.9 | 5.96 ± 0.57 | — | — | — | ||
| ► | H315N | 0.92 ± 0.26 | 1.35 ± 0.25 | 494.2 ± 20.3 | 393.33 ± 86.03 | 40.8 ± 0.3 | — | -1.5 | |
| ► | H315P | 1.04 ± 0.19 | 3.94 ± 0.38 | 433.4 ± 9.5 | 104.23 ± 12.82 | 38.8 ± 0.2 | 43.1 ± 0.0 | -1.6 | |
| H315Q | 0.41 | 1.12 ± 0.78 | 6.6 ± 1.1 | 5.88 ± 4.26 | 40.4 ± 0.5 | — | -1.1 | ||
| H315T | 4.76 | 4.67 ± 0.11 | 658.5 ± 4.0 | 141.04 ± 3.48 | — | — | — | ||
| H315Y | 0.90 | 7.02 ± 0.66 | 347.3 ± 9.4 | 49.47 ± 4.85 | — | 45.7 ± 0.1 | — | ||
| M316K | 0.35 | 3.77 ± 0.90 | 439.2 ± 25.9 | 116.43 ± 28.63 | — | — | — | ||
| E318R | 3.49 | 8.52 ± 0.97 | 5.3 ± 0.2 | 0.62 ± 0.07 | — | — | — | ||
| T321S | 1.84 | 9.44 ± 2.87 | 422.2 ± 42.1 | 44.72 ± 14.29 | — | 43.3 ± 0.4 | 2.4 | ||
| M323A | * | 9.34 ± 0.80 | 342.0 ± 8.7 | 36.59 ± 3.25 | 37.1 ± 0.1 | 42.5 ± 0.0 | — | ||
| M323C | 0.32 | 16.52 ± 1.78 | 105.5 ± 4.0 | 6.39 ± 0.73 | 39.9 ± 0.3 | — | 4.5 | ||
| M323D | 1.11 | 4.44 ± 0.89 | 5.4 ± 0.3 | 1.21 ± 0.25 | — | 40.3 ± 0.1 | 5.4 | ||
| ► | M323I | 5.16 ± 0.11 | 5.78 ± 0.33 | 327.0 ± 5.9 | 66.48 ± 6.22 | 36.1 ± 0.2 | 44.9 ± 0.1 | 6.4 | |
| M323K | — | — | — | — | — | — | — | ||
| ► | M323S | 0.17 ± 0.00 | 10.41 ± 1.33 | 503.0 ± 17.2 | 58.42 ± 5.44 | 38.6 ± 0.4 | — | 5.2 | |
| ► | W325A | 0.31 | 0.92 ± 0.20 | 52.2 ± 6.7 | 185.09 ± 179.05 | 36.4 ± 2,120.1 | 36.9 ± 1.1 | 7.0 | |
| W325C | * | 4.18 ± 0.53 | 8.7 ± 0.0 | 2.16 ± 0.22 | 39.3 ± 0.2 | 41.1 ± 0.0 | — | ||
| W325D | 2.22 | 27.09 ± 3.65 | 1,039.5 ± 55.4 | 38.38 ± 5.57 | — | — | — | ||
| W325E | 3.58 | 2.26 ± 0.28 | 1.6 ± 0.1 | 0.71 ± 0.09 | 38.0 ± 0.3 | — | 5.2 | ||
| W325F | 1.87 | 12.63 ± 3.27 | 116.0 ± 9.7 | 9.19 ± 2.50 | — | — | — | ||
| ► | W325G | * | — | — | — | 39.0 ± 0.6 | — | 10.0 | |
| W325H | * | 3.08 ± 0.43 | 28.1 ± 0.0 | 9.09 ± 1.30 | 37.7 ± 0.3 | 40.7 ± 0.0 | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| W325I | 3.45 | 1.29 ± 0.16 | 6.2 ± 0.2 | 4.84 ± 0.61 | 39.6 ± 0.5 | — | 29.7 | ||
| ► | W325K | 0.74 ± 0.45 | 3.38 ± 0.88 | 5.9 ± 0.4 | 1.61 ± 0.38 | 39.4 ± 0.4 | — | 3.6 | |
| W325L | * | 5.74 ± 0.35 | 88.8 ± 2.2 | 15.37 ± 1.08 | 40.4 ± 0.5 | — | — | ||
| ► | W325M | 3.80 ± 2.51 | 5.64 ± 2.32 | 39.1 ± 3.1 | 6.86 ± 1.91 | 38.8 ± 0.1 | 43.8 ± 0.3 | 3.5 | |
| W325N | 3.86 | 1.93 ± 0.15 | 4.4 ± 0.1 | 2.27 ± 0.18 | 40.4 ± 0.4 | — | 6.2 | ||
| W325P | — | — | — | — | — | — | 112.0 | ||
| ► | W325Q | 0.89 ± 0.87 | 0.60 ± 0.16 | 43.9 ± 2.8 | 121.93 ± 47.54 | 39.7 ± 0.2 | 42.2 ± 0.0 | 3.9 | |
| ► | W325R | 2.05 | 14.18 ± 3.12 | 213.4 ± 15.7 | 15.05 ± 3.49 | 40.0 ± 0.2 | 42.7 ± 0.0 | 3.0 | |
| W325S | 1.89 | 1.05 ± 0.21 | 6.1 ± 0.2 | 5.81 ± 1.17 | 42.0 ± 1.5 | — | 7.2 | ||
| W325T | 3.96 | 2.03 ± 0.26 | 7.0 ± 0.2 | 3.45 ± 0.46 | 38.2 ± 0.5 | — | 8.9 | ||
| ► | W325V | 2.59 ± 0.91 | 1.43 ± 0.16 | 8.6 ± 0.2 | 6.04 ± 0.71 | 39.1 ± 0.7 | — | 41.5 | |
| ► | W325Y | 3.02 | 2.41 ± 0.20 | 32.6 ± 0.6 | 13.52 ± 1.16 | 40.8 ± 0.2 | — | 5.1 | |
| H328A | 0.77 | 8.24 ± 0.32 | 423.2 ± 4.9 | 51.36 ± 2.09 | — | 45.4 ± 0.1 | — | ||
| ► | H328K | 1.83 ± 0.26 | 6.18 ± 2.38 | 306.7 ± 35.1 | 81.33 ± 27.49 | 39.0 ± 0.3 | — | 1.1 | |
| H328P | 0.36 | 2.71 ± 0.67 | 4.3 ± 0.2 | 1.58 ± 0.40 | — | — | 51.0 | ||
| P329N | — | — | — | — | — | — | — | ||
| S331A | * | 4.34 ± 0.11 | 668.9 ± 4.3 | 154.13 ± 4.11 | 39.3 ± 0.3 | 41.0 ± 0.0 | — | ||
| S331G | 2.43 | 5.82 ± 0.16 | 1,425.9 ± 14.4 | 244.86 ± 7.02 | — | — | — | ||
| S331T | 0.89 | 4.50 ± 1.38 | 27.4 ± 1.9 | 6.09 ± 1.91 | — | — | — | ||
| ► | Y333F | 2.69 ± 0.73 | 2.97 ± 0.37 | 338.2 ± 12.1 | 160.14 ± 31.23 | 37.8 ± 59,885.7 | 43.4 ± 0.3 | 0.3 | |
| K334D | 1.20 | 6.11 ± 0.90 | 279.9 ± 11.1 | 45.78 ± 6.95 | — | — | — | ||
| L336S | 0.28 | 8.02 ± 0.47 | 189.2 ± 3.1 | 23.58 ± 1.43 | — | — | — | ||
| ► | E340A | 2.37 ± 1.01 | 2.04 ± 0.29 | 361.8 ± 13.3 | 238.14 ± 90.24 | 38.3 ± 0.2 | 45.1 ± 0.1 | -3.2 | |
| E340S | 0.81 | 3.54 ± 0.46 | 308.7 ± 10.1 | 87.21 ± 11.72 | — | 45.0 ± 0.3 | 4.4 | ||
| ► | K341A | 3.14 | 5.33 ± 0.37 | 610.5 ± 10.6 | 113.48 ± 7.96 | 40.0 ± 0.1 | 45.1 ± 0.5 | 2.9 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | F343E | 0.10 | 45.41 ± 19.90 | 29.0 ± 6.2 | 0.64 ± 0.31 | — | — | -3.1 | |
| F343S | — | — | — | — | — | — | — | ||
| ► | L350I | 2.54 ± 0.87 | 4.71 ± 0.43 | 281.6 ± 8.4 | 50.60 ± 5.75 | — | 47.7 ± 0.1 | 0.1 | |
| I351V | 3.95 | 4.78 ± 0.39 | 436.1 ± 9.2 | 91.20 ± 7.74 | — | — | — | ||
| ► | T352A | 0.47 ± 0.43 | 11.50 ± 1.30 | 83.1 ± 2.1 | 8.39 ± 0.65 | 38.1 ± 0.3 | 40.2 ± 0.4 | 1.9 | |
| T352G | 0.43 | 6.83 ± 0.37 | 21.1 ± 0.4 | 3.09 ± 0.18 | — | — | 4.4 | ||
| T352P | 0.24 | 5.21 ± 0.36 | 252.6 ± 4.6 | 48.50 ± 3.42 | — | — | — | ||
| ► | T352S | 0.23 | 5.11 ± 1.38 | 234.4 ± 18.2 | 45.83 ± 12.86 | 37.5 ± 0.3 | — | — | |
| ► | T352V | 0.05 ± 0.01 | 2.68 ± 0.45 | 79.1 ± 3.1 | 29.52 ± 5.06 | — | — | — | |
| T352W | — | — | — | — | — | — | 71.5 | ||
| E353A | * | — | — | — | — | — | — | ||
| E353D | 0.45 | 2.35 ± 0.99 | 1.6 ± 0.2 | 0.68 ± 0.29 | — | — | — | ||
| N354A | * | 5.38 ± 0.67 | 2.2 ± 0.0 | 0.43 ± 0.00 | 40.8 ± 0.4 | 46.7 ± 0.0 | — | ||
| ► | N354S | 0.63 ± 0.38 | 3.34 ± 0.44 | 54.6 ± 2.2 | 20.46 ± 4.29 | — | 40.9 ± 0.3 | 1.8 | |
| G355A | — | — | — | — | — | — | — | ||
| G355N | 0.31 | 8.65 ± 2.96 | 0.4 ± 0.0 | 0.04 ± 0.01 | — | — | -3.6 | ||
| A356T | 1.90 | 2.89 ± 0.18 | 285.4 ± 4.2 | 98.63 ± 6.22 | — | 45.1 ± 0.1 | — | ||
| M358T | * | 4.83 ± 0.48 | 357.2 ± 8.7 | 74.04 ± 7.58 | 39.5 ± 0.2 | — | — | ||
| L362M | * | 5.20 ± 0.19 | 751.2 ± 6.5 | 144.39 ± 5.41 | 40.0 ± 0.2 | — | — | ||
| V363N | 2.07 | 4.64 ± 0.32 | 478.2 ± 8.3 | 103.16 ± 7.24 | — | — | — | ||
| ► | V363Q | 1.80 ± 0.67 | 5.67 ± 0.41 | 202.7 ± 6.4 | 36.86 ± 4.52 | — | 42.0 ± 0.4 | 1.6 | |
| G365N | 3.39 | 4.77 ± 0.12 | 961.6 ± 8.5 | 201.64 ± 5.33 | — | — | — | ||
| R372F | 0.41 | 4.84 ± 1.70 | 1.0 ± 0.1 | 0.21 ± 0.08 | — | — | 5.8 | ||
| R372H | 3.57 | 3.73 ± 0.33 | 625.2 ± 14.0 | 167.65 ± 15.29 | — | — | 2.9 | ||
| R372W | 0.51 | 0.56 ± 0.22 | 0.3 ± 0.0 | 0.56 ± 0.22 | — | — | 9.0 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| R372Y | 0.42 | 2.88 ± 0.47 | 1.7 ± 0.1 | 0.58 ± 0.10 | — | — | 6.8 | ||
| H373R | * | 6.31 ± 0.30 | 580.2 ± 6.5 | 92.00 ± 4.55 | 39.2 ± 0.3 | 51.4 ± 0.0 | — | ||
| Y375H | 0.49 | 5.45 ± 0.36 | 418.7 ± 7.4 | 76.84 ± 5.30 | — | — | — | ||
| Y375T | 0.43 | — | — | 0.09 ± 0.00 | — | — | 8.2 | ||
| E377Q | 0.31 | 4.86 ± 0.69 | 1,537.4 ± 58.0 | 316.60 ± 46.56 | 39.6 ± 0.2 | — | — | ||
| E377S | 1.11 | 4.64 ± 0.69 | 590.6 ± 22.4 | 127.35 ± 19.44 | — | — | — | ||
| H379R | * | — | — | — | — | — | — | ||
| H379T | — | — | — | — | — | — | — | ||
| ► | H379Y | 0.61 ± 0.25 | 2.52 ± 0.99 | 200.8 ± 18.9 | 628.48 ± 1,425.52 | 39.3 ± 0.3 | — | -0.9 | |
| H384A | 1.09 | 6.22 ± 0.42 | 644.9 ± 11.9 | 103.69 ± 7.30 | — | — | — | ||
| ► | H384I | 3.42 ± 0.47 | 2.73 ± 0.31 | 183.2 ± 5.2 | 68.95 ± 8.16 | — | 40.2 ± 0.2 | 12.9 | |
| H384R | 0.43 | 6.00 ± 0.39 | 31.4 ± 0.5 | 5.24 ± 0.35 | — | — | — | ||
| I387K | 0.17 | 9.29 ± 1.08 | 94.5 ± 3.4 | 10.18 ± 1.24 | — | 43.6 ± 0.4 | 4.7 | ||
| E388F | 3.62 | 4.79 ± 0.37 | 317.7 ± 6.5 | 66.28 ± 5.31 | — | — | — | ||
| E388G | 5.11 | 1.62 ± 0.21 | 177.9 ± 5.0 | 109.58 ± 14.28 | — | 44.6 ± 0.1 | -4.4 | ||
| ► | E388H | 0.77 ± 0.03 | 3.94 ± 0.60 | 245.5 ± 17.2 | 111.21 ± 19.28 | 40.1 ± 0.2 | — | -3.3 | |
| ► | E388L | 3.04 ± 0.66 | 3.64 ± 0.73 | 113.2 ± 4.1 | 87.02 ± 38.90 | 40.2 ± 0.2 | 44.9 ± 0.4 | -0.4 | |
| E388R | 4.14 | 3.67 ± 0.80 | 304.6 ± 16.4 | 82.96 ± 18.62 | — | — | — | ||
| E389R | * | — | — | — | — | 45.2 ± 0.1 | -0.7 | ||
| Q392N | 0.47 | 4.84 ± 1.10 | 331.4 ± 21.1 | 68.41 ± 16.07 | — | — | — | ||
| Y396C | — | — | — | — | — | — | 0.9 | ||
| Y396D | — | — | — | — | — | — | 9.4 | ||
| Y396E | — | — | — | — | — | — | 11.3 | ||
| Y396F | 1.14 | 6.40 ± 1.03 | 155.0 ± 7.0 | 24.23 ± 4.03 | 38.0 ± 0.2 | — | 1.8 | ||
| Y396G | — | — | — | — | — | — | 14.6 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | Y396P | — | — | — | — | — | — | 283.8 | |
| Y396Q | — | — | — | — | — | — | 9.0 | ||
| Y396R | — | — | — | — | — | — | 9.3 | ||
| Y396S | — | — | — | — | — | — | 8.2 | ||
| Y396V | — | — | — | — | — | — | 21.5 | ||
| Y396W | 1.28 | 1.96 ± 0.54 | 14.1 ± 1.1 | 7.20 ± 2.05 | 38.4 ± 0.5 | — | 3.9 | ||
| F397A | 0.68 | 2.78 ± 0.61 | 25.1 ± 1.4 | 9.03 ± 2.03 | 39.2 ± 4.4 | 39.1 ± 2.9 | 6.7 | ||
| F397D | — | — | — | — | — | — | 7.9 | ||
| F397E | — | — | — | — | — | — | 0.0 | ||
| F397G | — | — | — | — | — | — | 12.0 | ||
| F397H | — | — | — | — | — | — | 3.8 | ||
| F397I | 2.20 | 2.02 ± 0.28 | 79.4 ± 2.5 | 39.24 ± 5.56 | 37.5 ± 0.1 | 37.2 ± 0.8 | 26.9 | ||
| F397K | — | — | — | — | — | — | 0.0 | ||
| F397N | 0.71 | 8.68 ± 0.95 | 86.3 ± 3.1 | 9.94 ± 1.14 | 38.5 ± 0.8 | 38.5 ± 0.8 | 6.0 | ||
| F397R | — | — | — | — | — | — | 8.6 | ||
| F397S | 1.10 | 4.81 ± 0.23 | 88.0 ± 1.1 | 18.31 ± 0.91 | 35.7 ± 0.7 | 35.7 ± 0.7 | 7.6 | ||
| F397V | 0.50 | 3.01 ± 0.44 | 7.1 ± 0.2 | 2.38 ± 0.36 | 36.7 ± 0.3 | — | 27.7 | ||
| F397W | 1.37 | 9.02 ± 0.79 | 105.0 ± 3.3 | 11.64 ± 1.09 | 38.2 ± 0.4 | 38.2 ± 0.4 | 0.5 | ||
| F397Y | — | — | — | — | — | — | 1.6 | ||
| V398N | 0.41 | 4.11 ± 0.25 | 400.5 ± 6.3 | 97.37 ± 6.16 | 34.3 ± 0.2 | — | 4.6 | ||
| W399A | * | — | — | — | — | — | — | ||
| W399C | * | 70.33 ± 5.89 | 2.2 ± 0.0 | — | 41.7 ± 0.1 | 44.8 ± 0.0 | — | ||
| W399G | * | — | — | — | — | — | — | ||
| W399Q | 3.85 | 21.84 ± 3.20 | 0.6 ± 0.0 | 0.03 ± 0.00 | — | 44.8 ± 0.1 | 6.4 | ||
| W399R | * | 12.75 ± 0.87 | — | — | 37.9 ± 0.8 | 49.1 ± 0.0 | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| W399S | * | — | — | — | — | — | — | ||
| S400A | * | 3.22 ± 0.22 | 435.1 ± 6.5 | 135.09 ± 9.53 | 39.9 ± 0.3 | 47.3 ± 0.0 | — | ||
| S400G | 2.18 | 30.88 ± 2.17 | 287.8 ± 8.3 | 9.32 ± 0.71 | — | — | — | ||
| S400W | — | — | — | — | — | — | 124.8 | ||
| F401K | 0.63 | — | — | — | — | 44.9 ± 0.8 | 18.4 | ||
| L402C | 5.89 | 3.12 ± 0.21 | 744.3 ± 24.0 | 238.68 ± 17.68 | — | — | — | ||
| L402E | 0.27 | 2.58 ± 0.14 | 43.7 ± 0.6 | 16.96 ± 0.95 | — | — | — | ||
| D403A | — | — | — | — | — | — | — | ||
| ► | N404A | 3.83 | 7.75 ± 1.29 | 2.8 ± 0.1 | 0.32 ± 0.04 | 41.9 ± 0.1 | 54.3 ± 0.0 | — | |
| ► | N404C | 1.55 | 18.77 ± 3.31 | 78.0 ± 4.5 | 4.25 ± 0.70 | 43.2 ± 0.2 | 46.0 ± 0.0 | -1.9 | |
| ► | N404D | 1.44 ± 0.25 | 27.12 ± 2.51 | 284.8 ± 14.9 | 62.38 ± 13.36 | — | 42.4 ± 1.8 | 5.7 | |
| ► | N404E | 1.75 ± 1.46 | 5.04 ± 0.94 | 53.5 ± 3.2 | 15.11 ± 3.71 | 40.5 ± 3.0 | 45.3 ± 0.6 | 4.2 | |
| ► | N404L | 1.90 ± 0.75 | 30.74 ± 2.08 | 22.8 ± 0.6 | 0.74 ± 0.05 | — | 44.6 ± 0.1 | -1.6 | |
| ► | N404M | 2.07 ± 0.94 | 19.36 ± 2.27 | 15.5 ± 1.4 | 1.29 ± 0.46 | 40.6 ± 0.3 | 46.6 ± 0.2 | -3.2 | |
| ► | N404Q | 2.24 ± 1.29 | 2.48 ± 0.81 | 0.2 ± 0.0 | 0.14 ± 0.12 | — | 48.3 ± 0.3 | 0.5 | |
| ► | N404S | 2.86 ± 2.10 | 26.79 ± 4.28 | 35.2 ± 1.7 | 1.28 ± 0.20 | 44.5 ± 0.1 | — | — | |
| E406A | * | — | — | — | 40.1 ± 0.7 | 49.0 ± 0.0 | — | ||
| ► | E406D | 0.49 | 29.16 ± 4.17 | 28.4 ± 1.5 | 0.94 ± 0.17 | 40.4 ± 0.2 | 42.2 ± 0.0 | -4.4 | |
| E406F | 0.64 | — | — | — | — | — | — | ||
| E406H | 2.36 | 0.02 ± 0.04 | 0.0 ± 0.0 | 1.80 ± 2.83 | — | — | 4.5 | ||
| E406K | * | — | — | — | — | 44.1 ± 0.3 | 25.2 | ||
| ► | E406Q | 3.81 ± 1.18 | 58.18 ± 5.12 | 36.9 ± 3.0 | 0.37 ± 0.07 | — | 47.2 ± 1.3 | 4.3 | |
| E406T | 3.40 | 2.57 ± 1.38 | 15.6 ± 1.9 | 6.05 ± 3.33 | — | — | — | ||
| ► | W407A | 0.91 | 27.09 ± 4.32 | 0.6 ± 0.0 | 0.02 ± 0.00 | — | 42.4 ± 0.2 | — | |
| ► | W407D | 0.57 ± 0.30 | 3.73 ± 3.39 | 1.6 ± 1.0 | 2.69 ± 11.66 | — | 43.3 ± 0.6 | -0.4 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| W407E | — | — | — | — | — | — | 10.6 | ||
| ► | W407F | 1.80 ± 0.64 | 33.76 ± 3.39 | 6.0 ± 0.2 | 0.18 ± 0.02 | 39.2 ± 0.6 | 43.8 ± 0.1 | 5.1 | |
| W407G | — | — | — | — | — | — | — | ||
| ► | W407H | 0.97 ± 0.47 | 5.58 ± 3.04 | 0.5 ± 0.1 | 1.02 ± 0.79 | — | — | — | |
| ► | W407K | 0.09 | 17.69 ± 3.06 | 3.6 ± 0.2 | 0.20 ± 0.04 | — | — | 3.5 | |
| W407L | * | — | — | — | — | — | 29.5 | ||
| ► | W407N | 0.20 ± 0.05 | — | — | — | — | — | -0.9 | |
| W407Q | — | — | — | — | — | — | — | ||
| ► | W407R | — | — | — | — | — | — | 8.8 | |
| W407S | — | — | — | — | — | — | -1.8 | ||
| W407Y | — | — | — | — | — | — | — | ||
| A408F | 0.31 | 37.97 ± 8.36 | 341.9 ± 33.2 | 9.00 ± 2.17 | — | — | — | ||
| ► | A408H | 1.16 ± 0.57 | 13.31 ± 0.87 | 258.9 ± 5.6 | 19.45 ± 1.36 | — | 43.7 ± 0.1 | 3.0 | |
| ► | A408K | 1.95 ± 1.11 | 5.34 ± 0.67 | 478.2 ± 16.0 | 89.76 ± 10.92 | — | 46.4 ± 0.2 | — | |
| A408S | 0.23 | 9.32 ± 2.00 | 158.5 ± 10.3 | 17.02 ± 3.81 | 38.4 ± 0.2 | — | 1.9 | ||
| A408T | 0.14 | 12.90 ± 2.77 | 153.9 ± 10.7 | 11.93 ± 2.69 | 37.3 ± 0.5 | — | 4.9 | ||
| ► | W409A | 1.93 ± 1.93 | 7.15 ± 1.40 | 184.0 ± 10.8 | 25.77 ± 4.75 | 40.2 ± 0.3 | — | 4.0 | |
| W409C | 6.00 | 2.44 ± 0.13 | 156.2 ± 1.9 | 64.01 ± 3.39 | 39.4 ± 2.2 | — | 5.0 | ||
| ► | W409D | 1.48 ± 1.51 | 25.96 ± 7.34 | 592.0 ± 31.6 | 182.58 ± 18.18 | 40.3 ± 0.3 | — | 6.8 | |
| ► | W409E | 1.72 ± 0.78 | 2.05 ± 0.21 | 462.4 ± 8.4 | 221.06 ± 15.66 | 39.6 ± 0.4 | 44.7 ± 0.1 | 4.9 | |
| ► | W409F | 0.62 ± 0.16 | 3.45 ± 0.25 | 605.1 ± 11.0 | 175.69 ± 13.06 | 39.6 ± 0.2 | — | 1.2 | |
| W409G | 0.44 | 3.85 ± 0.82 | 14.1 ± 0.7 | 3.66 ± 0.81 | — | — | — | ||
| ► | W409H | 1.39 ± 1.05 | 2.70 ± 0.26 | 375.1 ± 11.0 | 136.73 ± 18.14 | 38.9 ± 0.6 | — | 2.1 | |
| ► | W409I | 1.13 ± 0.00 | 6.88 ± 1.32 | 375.4 ± 31.9 | 41.51 ± 11.89 | 40.3 ± 0.3 | — | 8.2 | |
| W409K | 0.48 | 1.62 ± 0.22 | 180.6 ± 5.4 | 111.29 ± 15.53 | 38.5 ± 0.3 | — | 3.5 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| W409L | 6.10 | 2.05 ± 0.16 | 87.5 ± 1.6 | 42.57 ± 3.43 | 39.5 ± 0.3 | — | 9.5 | ||
| W409M | 0.62 | 7.11 ± 2.14 | 35.4 ± 2.6 | 4.98 ± 1.55 | 40.0 ± 0.1 | — | -0.6 | ||
| ► | W409N | 6.60 ± 1.67 | 5.91 ± 0.89 | 184.0 ± 8.7 | 27.96 ± 4.28 | 39.1 ± 0.5 | 38.9 ± 0.1 | 1.9 | |
| W409P | — | — | — | — | — | — | 82.7 | ||
| ► | W409R | 2.03 ± 1.08 | 2.48 ± 0.27 | 119.9 ± 3.4 | 47.31 ± 5.33 | 34.6 ± 0.6 | 44.5 ± 0.3 | 3.7 | |
| W409S | 6.17 | 4.52 ± 0.76 | 143.7 ± 6.3 | 31.79 ± 5.51 | 39.4 ± 0.4 | — | 4.1 | ||
| ► | W409T | 0.95 ± 0.83 | 3.68 ± 0.85 | 214.6 ± 14.1 | 90.81 ± 29.37 | 36.4 ± 1.3 | — | -356.9 | |
| W409V | 4.17 | 9.62 ± 1.08 | 139.5 ± 4.8 | 14.50 ± 1.71 | 39.3 ± 0.9 | — | 5.0 | ||
| ► | W409Y | 0.85 ± 0.59 | 2.97 ± 0.27 | 633.7 ± 17.3 | 217.18 ± 21.58 | 38.4 ± 0.3 | 43.5 ± 0.0 | 0.4 | |
| ► | Y411R | 0.28 ± 0.13 | 1.99 ± 0.63 | 50.3 ± 3.3 | 32.27 ± 11.15 | — | — | — | |
| K413A | 3.15 | 7.32 ± 0.60 | 381.9 ± 9.0 | 52.18 ± 4.42 | — | 48.8 ± 0.1 | — | ||
| K413Q | 3.02 | 5.41 ± 0.62 | 386.6 ± 12.1 | 71.53 ± 8.55 | — | 45.8 ± 1.7 | 5.0 | ||
| K413R | 0.41 | 0.85 ± 0.29 | 470.8 ± 32.0 | 555.04 ± 196.18 | 40.1 ± 0.1 | — | — | ||
| R414K | 0.55 | 26.88 ± 10.87 | 399.9 ± 66.3 | 14.88 ± 6.50 | 40.6 ± 0.6 | — | 1.6 | ||
| F415A | * | 16.63 ± 4.00 | — | — | 40.5 ± 0.3 | 44.7 ± 0.0 | — | ||
| F415H | 0.47 | 0.78 ± 0.34 | 9.2 ± 1.0 | 11.81 ± 5.23 | — | — | 2.3 | ||
| F415N | * | — | — | — | — | — | — | ||
| ► | F415Y | 2.11 ± 0.13 | 9.03 ± 0.99 | 202.3 ± 7.4 | 21.64 ± 2.64 | — | 41.6 ± 0.2 | — | |
| I417H | 1.25 | 2.81 ± 0.87 | 4.2 ± 0.3 | 1.51 ± 0.48 | — | 44.5 ± 0.8 | 8.6 | ||
| ► | H419Y | 3.27 ± 0.04 | 4.88 ± 0.44 | 742.6 ± 14.1 | 143.16 ± 11.99 | — | 49.1 ± 1.1 | -2.1 | |
| ► | N421D | 0.79 ± 0.24 | 3.26 ± 0.33 | 1,113.3 ± 26.0 | 356.21 ± 42.67 | 41.3 ± 0.2 | — | -0.2 | |
| Y422F | 2.83 | 30.66 ± 5.28 | 183.8 ± 13.5 | 6.00 ± 1.12 | — | 47.4 ± 0.2 | -0.4 | ||
| E423S | * | 6.60 ± 0.42 | 530.4 ± 34.6 | 80.31 ± 7.36 | 39.5 ± 0.1 | 44.8 ± 0.0 | — | ||
| T424H | 3.64 | 6.10 ± 0.34 | 39.6 ± 0.6 | 6.50 ± 0.37 | — | 46.7 ± 0.2 | 1.7 | ||
| ► | R427H | 0.73 ± 0.27 | 2.48 ± 0.47 | 4.1 ± 0.2 | 1.38 ± 0.27 | — | 44.3 ± 0.3 | 11.8 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | R427L | 1.00 ± 0.85 | 4.71 ± 0.38 | 377.3 ± 8.0 | 80.49 ± 6.64 | — | 44.7 ± 0.2 | 4.9 | |
| R427W | 0.31 | 0.98 ± 0.27 | 35.6 ± 2.0 | 36.11 ± 10.00 | 40.2 ± 0.1 | — | — | ||
| P429A | 2.51 | 9.81 ± 1.03 | 327.2 ± 10.7 | 33.35 ± 3.65 | — | — | — | ||
| ► | L434S | 1.73 ± 0.76 | 1.69 ± 0.18 | 221.9 ± 9.6 | 244.50 ± 85.67 | — | 43.0 ± 0.0 | 1.2 | |
| ► | W435Y | 0.48 | 7.41 ± 0.72 | 823.9 ± 23.2 | 111.17 ± 11.22 | 40.2 ± 0.2 | — | — | |
| A441V | 2.07 | 4.65 ± 0.10 | 859.5 ± 4.6 | 184.98 ± 4.01 | — | — | — |