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 | — | |
| W14F | 0.71 | 5.33 ± 0.40 | 836.9 ± 16.4 | 156.97 ± 12.12 | — | — | — | ||
| G15N | — | — | — | — | — | — | — | ||
| ► | T16K | 0.69 | 0.95 ± 0.39 | 0.3 ± 0.0 | 0.36 ± 0.15 | — | 46.2 ± 2.1 | 36.0 | |
| ► | T16S | 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 | |
| ► | S17A | 1.69 | 5.57 ± 0.80 | 219.7 ± 7.9 | 46.68 ± 7.92 | 40.2 ± 0.3 | 43.8 ± 0.1 | 1.8 | |
| ► | T18A | * | 5.30 ± 0.48 | 504.4 ± 13.0 | 95.25 ± 8.88 | 38.2 ± 0.4 | 42.0 ± 0.0 | 9.3 | |
| S19A | * | 14.01 ± 0.40 | 125.6 ± 0.0 | 8.88 ± 0.22 | 39.4 ± 0.3 | 43.9 ± 0.0 | — | ||
| S19N | — | — | — | — | — | — | — | ||
| ► | S20A | 3.96 | 12.48 ± 2.65 | 445.8 ± 44.5 | 33.95 ± 6.06 | 40.2 ± 694,236.0 | — | — | |
| S20E | * | 7.32 ± 0.38 | 526.1 ± 6.5 | 71.87 ± 3.90 | 39.5 ± 0.1 | — | — | ||
| Y21A | * | 31.55 ± 3.61 | 162.4 ± 6.5 | 5.20 ± 0.65 | 38.3 ± 0.2 | 40.6 ± 0.0 | — | ||
| ► | Q22A | 1.25 | 1.68 ± 1.14 | 0.1 ± 0.0 | 0.08 ± 0.05 | 35.2 ± 0.7 | — | -1.3 | |
| Q22C | * | — | — | — | — | — | — | ||
| Q22D | 0.57 | — | — | — | — | — | — | ||
| ► | Q22M | 0.43 | — | — | — | — | 46.4 ± 1.5 | -18.7 | |
| Q22P | — | — | — | — | — | — | — | ||
| Q22S | * | — | — | — | 36.8 ± 0.2 | 43.1 ± 0.0 | — | ||
| ► | Q22T | 1.54 ± 0.65 | 3.37 ± 0.64 | 120.1 ± 3.1 | 24.01 ± 2.42 | — | 42.8 ± 0.0 | -0.3 | |
| I23N | 0.31 | 19.41 ± 4.87 | 44.5 ± 4.0 | 2.29 ± 0.61 | — | — | — | ||
| E24C | — | — | — | — | — | — | 5.4 | ||
| E24D | 0.41 | 4.55 ± 0.53 | 32.0 ± 1.0 | 7.04 ± 0.86 | 37.5 ± 0.3 | 37.5 ± 0.3 | 4.4 | ||
| E24F | — | — | — | — | — | — | 32.3 | ||
| E24G | — | — | — | — | — | — | 8.0 | ||
| E24H | — | — | — | — | — | — | 22.3 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| E24I | — | — | — | — | — | — | 32.6 | ||
| E24K | — | — | — | — | — | — | 27.7 | ||
| E24N | — | — | — | — | — | — | 6.4 | ||
| E24R | — | — | — | — | — | — | 16.9 | ||
| E24T | — | — | — | — | — | — | 6.5 | ||
| E24W | — | — | — | — | — | — | 383.6 | ||
| E24Y | 4.45 | 1.52 ± 0.20 | 184.4 ± 5.4 | 120.95 ± 16.52 | 40.3 ± 0.3 | 40.3 ± 0.3 | 27.1 | ||
| ► | G26A | 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 | |
| ► | D28Q | 3.48 ± 2.87 | 3.45 ± 0.63 | 307.9 ± 13.3 | 87.18 ± 12.62 | 40.3 ± 0.1 | 45.5 ± 0.0 | — | |
| E29K | 0.62 | 6.81 ± 0.48 | 638.8 ± 12.9 | 93.81 ± 6.93 | 38.4 ± 0.3 | — | -0.9 | ||
| S35L | — | — | — | — | — | — | — | ||
| W37A | — | — | — | — | — | — | — | ||
| D38E | 2.00 | 2.72 ± 0.31 | 16.3 ± 0.4 | 5.97 ± 0.70 | — | — | 11.6 | ||
| ► | D38W | 0.40 ± 0.06 | 2.62 ± 0.82 | 7.5 ± 0.5 | 2.74 ± 0.71 | — | 46.8 ± 3.6 | 14.3 | |
| T39I | — | — | — | — | — | — | — | ||
| T39N | 5.49 | 6.57 ± 1.21 | 23.9 ± 1.2 | 3.63 ± 0.69 | — | 46.8 ± 0.1 | -0.2 | ||
| F40N | 0.26 | 4.88 ± 0.15 | 8.3 ± 0.1 | 1.70 ± 0.06 | — | — | — | ||
| ► | C41S | 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 | |
| I43T | 1.41 | 4.05 ± 0.21 | 265.2 ± 3.8 | 65.54 ± 3.50 | — | 43.3 ± 0.2 | 2.2 | ||
| P44L | * | — | — | — | — | — | — | ||
| G49C | 0.04 | 40.81 ± 25.63 | 4.4 ± 1.2 | 0.11 ± 0.07 | — | — | — | ||
| C52S | 0.85 | 6.63 ± 3.19 | 570.0 ± 74.3 | 86.02 ± 42.89 | 40.0 ± 0.0 | — | — | ||
| V55A | 2.06 | 5.30 ± 0.91 | 107.2 ± 5.0 | 20.22 ± 3.61 | — | 40.8 ± 0.2 | — | ||
| V55G | * | 8.25 ± 0.54 | 562.9 ± 10.8 | 68.19 ± 4.55 | 39.3 ± 0.1 | — | — | ||
| ► | F60Y | 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 |
||
| H61R | 1.22 | 5.64 ± 0.68 | 665.5 ± 21.2 | 118.04 ± 14.68 | — | — | — | ||
| ► | H62R | 2.55 ± 1.58 | 5.90 ± 0.52 | 302.3 ± 6.9 | 74.39 ± 6.46 | — | 49.8 ± 0.3 | -0.2 | |
| K64A | 1.55 | 3.79 ± 0.40 | 167.6 ± 4.4 | 44.28 ± 4.78 | 38.7 ± 0.4 | — | 1.3 | ||
| ► | K64C | 2.86 ± 0.01 | 1.75 ± 0.23 | 115.8 ± 3.3 | 67.22 ± 8.78 | 38.1 ± 0.3 | — | 2.4 | |
| K64D | 1.11 | 3.96 ± 0.39 | 116.3 ± 2.8 | 29.36 ± 2.96 | 38.3 ± 0.3 | — | 5.6 | ||
| K64E | 1.36 | 3.79 ± 0.28 | 141.6 ± 2.7 | 37.39 ± 2.87 | 38.8 ± 0.2 | — | 0.8 | ||
| ► | K64F | 2.58 ± 0.71 | 3.29 ± 0.25 | 70.9 ± 1.6 | 23.62 ± 2.25 | 37.4 ± 0.5 | — | 7.6 | |
| ► | K64G | 1.36 ± 0.45 | 3.02 ± 0.21 | 102.4 ± 2.0 | 26.48 ± 2.04 | 38.0 ± 0.5 | — | 4.6 | |
| K64H | 6.16 | 1.95 ± 0.19 | 164.0 ± 3.6 | 84.15 ± 8.28 | 38.2 ± 0.1 | — | 3.3 | ||
| K64I | 1.80 | 2.43 ± 0.19 | 104.7 ± 2.0 | 43.13 ± 3.46 | 38.4 ± 0.2 | — | 1.1 | ||
| K64L | 0.83 | 2.37 ± 0.30 | 60.7 ± 1.9 | 25.68 ± 3.33 | 40.6 ± 0.8 | — | 5.5 | ||
| K64M | 4.60 | 4.59 ± 0.53 | 200.1 ± 6.0 | 43.57 ± 5.17 | 38.0 ± 0.1 | — | 2.6 | ||
| ► | K64N | 3.50 ± 0.22 | 4.16 ± 0.41 | 156.9 ± 4.4 | 37.68 ± 4.19 | 35.6 ± 7.2 | — | 3.1 | |
| K64P | 1.49 | 2.90 ± 0.26 | 81.6 ± 1.8 | 28.19 ± 2.62 | 38.9 ± 0.3 | — | 11.7 | ||
| K64Q | 1.42 | 4.28 ± 1.38 | 137.0 ± 12.5 | 31.98 ± 10.68 | 39.3 ± 0.2 | — | 0.8 | ||
| K64R | 2.60 | 2.82 ± 0.25 | 109.8 ± 2.3 | 38.93 ± 3.50 | 39.2 ± 0.3 | — | 1.4 | ||
| ► | K64S | 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 | |
| K64W | 1.94 | 3.95 ± 0.43 | 74.2 ± 2.2 | 18.78 ± 2.13 | 38.2 ± 0.7 | — | 7.8 | ||
| K64Y | 0.47 | 1.86 ± 0.17 | 93.3 ± 1.9 | 50.18 ± 4.73 | 38.0 ± 0.5 | — | 7.7 | ||
| D66A | 0.01 | — | — | 0.33 ± 0.09 | — | — | 0.0 | ||
| D66C | 0.98 | 0.69 ± 1.31 | 0.0 ± 0.0 | 0.07 ± 0.14 | — | — | — | ||
| D66F | 0.77 | 4.13 ± 9.33 | 0.1 ± 0.0 | 0.01 ± 0.03 | — | — | — | ||
| ► | D66M | 0.36 ± 0.26 | 0.13 ± 0.81 | 0.1 ± 0.1 | 1.96 ± 14.10 | — | — | — | |
| D66N | 0.44 | 0.28 ± 0.43 | 0.2 ± 0.0 | 0.56 ± 0.87 | — | — | — | ||
| D66P | 0.28 | — | — | — | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| D66R | 0.13 | — | — | 0.02 ± 0.00 | — | — | — | ||
| D66S | 0.35 | 0.09 ± 0.16 | 0.1 ± 0.0 | 1.39 ± 2.61 | — | — | — | ||
| ► | K71H | 3.26 ± 0.47 | 4.02 ± 0.85 | 326.4 ± 14.3 | 82.25 ± 14.18 | — | 46.1 ± 0.8 | 2.4 | |
| F75A | * | 5.47 ± 0.28 | 502.2 ± 6.5 | 91.79 ± 4.76 | 38.6 ± 0.4 | — | — | ||
| ► | Y78F | 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 | |
| ► | Y78N | 0.15 ± 0.06 | 4.98 ± 0.47 | 364.4 ± 11.9 | 102.68 ± 19.80 | 38.0 ± 1.2 | — | 10.8 | |
| R79A | — | — | — | — | — | — | — | ||
| R79L | 0.54 | 43.36 ± 1.82 | 7.4 ± 0.1 | 0.17 ± 0.01 | — | — | — | ||
| R79Q | 0.70 | 6.62 ± 2.33 | 0.8 ± 0.1 | 0.13 ± 0.05 | — | — | -0.1 | ||
| F80D | 0.22 | — | — | — | — | — | — | ||
| F80W | 0.38 | 73.32 ± 94.87 | 20.0 ± 14.3 | 0.27 ± 0.40 | — | — | — | ||
| S81A | 3.12 | 20.08 ± 1.37 | 190.0 ± 4.9 | 9.46 ± 0.69 | — | 37.5 ± 0.1 | 1.9 | ||
| ► | V82I | 3.56 ± 0.08 | 3.21 ± 0.31 | 508.4 ± 11.5 | 158.50 ± 16.22 | 39.2 ± 0.2 | — | — | |
| V82T | 2.67 | 5.29 ± 2.61 | 239.4 ± 31.8 | 45.23 ± 23.12 | — | 46.7 ± 0.1 | 4.7 | ||
| ► | A83P | 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 | |
| ► | A83S | 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 | |
| ► | A91E | 2.47 ± 0.38 | 3.87 ± 0.60 | 317.7 ± 12.0 | 190.09 ± 62.25 | — | 45.7 ± 0.2 | -1.4 | |
| A91K | 0.79 | 4.46 ± 0.60 | 307.4 ± 10.5 | 68.93 ± 9.53 | — | — | — | ||
| I93V | 1.25 | 6.67 ± 0.28 | 790.7 ± 9.2 | 118.48 ± 5.19 | — | — | — | ||
| ► | I94A | 0.42 ± 0.31 | 7.44 ± 2.58 | 603.9 ± 21.6 | 109.73 ± 11.98 | 38.5 ± 304,961.7 | — | -2.8 | |
| I94C | 5.45 | 3.58 ± 0.15 | 19.6 ± 0.2 | 5.47 ± 0.24 | 38.1 ± 0.2 | — | — | ||
| I94D | 0.72 | 11.21 ± 0.69 | 89.6 ± 1.8 | 7.99 ± 0.51 | 37.4 ± 183,653.0 | — | — | ||
| I94E | * | 6.71 ± 0.79 | 692.7 ± 28.1 | 103.26 ± 12.77 | 39.7 ± 0.3 | — | — | ||
| I94G | 0.45 | 9.30 ± 3.31 | 93.7 ± 10.2 | 10.08 ± 3.75 | — | — | — | ||
| I94H | 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 |
||
| I94K | 0.31 | 3.73 ± 0.92 | 4,015.9 ± 262.1 | 1,075.53 ± 275.42 | 39.7 ± 0.3 | — | — | ||
| I94M | 0.87 | 9.04 ± 0.55 | 148.5 ± 2.8 | 16.43 ± 1.05 | 37.5 ± 0.3 | — | — | ||
| I94N | 1.50 | 9.39 ± 0.98 | 183.9 ± 5.9 | 19.59 ± 2.14 | 36.0 ± 0.2 | — | — | ||
| I94Q | 3.47 | 13.34 ± 0.58 | 209.0 ± 3.0 | 15.66 ± 0.72 | 37.8 ± 2,701,870.0 | — | — | ||
| ► | I94R | 0.53 ± 0.37 | 7.38 ± 1.79 | 239.8 ± 13.3 | 33.82 ± 5.66 | 37.6 ± 0.2 | — | -8.0 | |
| I94S | 1.40 | 10.13 ± 3.43 | 299.2 ± 29.0 | 29.54 ± 10.41 | 36.8 ± 0.2 | — | — | ||
| I94V | 1.88 | 10.44 ± 1.37 | 22.5 ± 0.9 | 2.16 ± 0.30 | 37.9 ± 88,473.6 | — | — | ||
| I94Y | 2.71 | 10.90 ± 0.60 | 248.8 ± 4.3 | 22.81 ± 1.31 | 35.8 ± 0.2 | — | — | ||
| H104R | * | 10.62 ± 0.53 | 868.1 ± 13.0 | 81.83 ± 4.33 | 40.1 ± 0.1 | 48.2 ± 0.0 | — | ||
| I109A | 1.03 | 5.15 ± 0.34 | 732.2 ± 13.0 | 142.12 ± 9.74 | — | — | — | ||
| ► | L111E | 2.77 ± 0.93 | 6.39 ± 0.81 | 387.5 ± 11.1 | 68.79 ± 5.53 | — | 43.3 ± 0.0 | -0.5 | |
| L111M | 2.35 | 4.14 ± 0.35 | 391.2 ± 8.6 | 94.54 ± 8.36 | — | 47.3 ± 0.1 | 0.6 | ||
| L111N | 2.91 | 4.27 ± 0.67 | 330.4 ± 13.4 | 77.38 ± 12.49 | — | 44.8 ± 0.0 | 0.0 | ||
| L111S | 0.26 | 2.76 ± 0.50 | 95.4 ± 4.1 | 34.55 ± 6.39 | 38.1 ± 0.5 | — | -1.1 | ||
| ► | M117S | 0.93 ± 0.25 | 9.80 ± 0.84 | 153.1 ± 4.9 | 13.53 ± 1.21 | — | 40.0 ± 34.1 | 5.0 | |
| ► | T119I | 1.99 ± 1.06 | 21.67 ± 8.57 | 310.5 ± 77.4 | 14.45 ± 10.02 | — | 43.5 ± 0.3 | 4.2 | |
| T119S | 1.61 | 2.21 ± 0.44 | 138.1 ± 6.3 | 62.58 ± 12.74 | — | 43.1 ± 0.1 | 4.0 | ||
| Y121A | 0.31 | 11.90 ± 1.18 | 11.2 ± 0.4 | 0.94 ± 0.10 | — | 46.0 ± 0.1 | — | ||
| ► | Y121F | 2.03 ± 1.69 | 0.87 ± 0.86 | 108.4 ± 20.7 | 124.90 ± 126.20 | — | 44.5 ± 0.6 | 3.2 | |
| H122A | * | 15.10 ± 3.36 | 116.9 ± 8.7 | 7.79 ± 1.73 | 40.2 ± 0.2 | — | — | ||
| ► | H122D | 0.39 ± 0.17 | — | — | 1.48 ± 0.23 | — | — | 3.9 | |
| H122E | — | — | — | — | — | — | — | ||
| H122M | 0.55 | 17.02 ± 4.07 | 9.8 ± 0.8 | 0.57 ± 0.15 | — | — | — | ||
| H122N | * | 23.22 ± 2.20 | 2.2 ± 0.0 | — | 40.3 ± 0.6 | — | — | ||
| H122Q | 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 |
||
| ► | H122T | 1.09 ± 0.25 | 66.09 ± 31.10 | 20.7 ± 5.2 | 0.28 ± 0.12 | — | 45.1 ± 0.5 | 3.1 | |
| H122V | 0.43 | 44.36 ± 5.88 | 5.2 ± 0.3 | 0.12 ± 0.02 | — | — | — | ||
| W123A | — | — | — | — | — | — | — | ||
| ► | W123D | 0.21 ± 0.03 | 13.94 ± 4.28 | 93.9 ± 12.6 | 4.31 ± 2.14 | 35.6 ± 0.0 | — | 5.9 | |
| ► | W123E | 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 | |
| W123F | * | 16.08 ± 2.07 | 387.5 ± 17.3 | 24.03 ± 3.25 | 41.7 ± 0.4 | 50.7 ± 0.0 | — | ||
| W123G | 0.38 | — | — | 0.12 ± 0.00 | — | — | — | ||
| W123H | * | 89.18 ± 4.31 | 69.3 ± 2.2 | 0.87 ± 0.00 | 40.1 ± 0.1 | 46.2 ± 0.0 | — | ||
| W123K | 0.25 | 56.50 ± 10.55 | 1.7 ± 0.2 | 0.03 ± 0.01 | — | — | — | ||
| W123L | 1.43 | — | — | 0.61 ± 0.02 | 37.6 ± 633,161.0 | — | — | ||
| W123M | 1.46 | 50.86 ± 3.20 | 100.2 ± 3.2 | 1.97 ± 0.14 | — | 43.9 ± 0.1 | 5.2 | ||
| ► | W123N | 0.59 ± 0.30 | 11.72 ± 8.19 | 3.0 ± 0.4 | 2.56 ± 1.49 | — | 48.2 ± 0.1 | 4.6 | |
| W123P | — | — | — | — | — | — | — | ||
| W123Q | 0.63 | — | — | 0.29 ± 0.02 | — | — | — | ||
| ► | W123S | 0.56 ± 0.18 | 13.51 ± 2.65 | 1.2 ± 0.1 | 0.10 ± 0.01 | 40.3 ± 0.9 | — | 0.0 | |
| W123T | 0.46 | 0.37 ± 0.25 | 0.4 ± 0.0 | 1.01 ± 0.69 | — | 47.0 ± 2.1 | -0.2 | ||
| ► | W123Y | 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 | |
| D124F | — | — | — | — | — | — | — | ||
| ► | L125Q | 0.44 ± 0.24 | 2.46 ± 0.60 | 450.5 ± 15.3 | 169.36 ± 21.40 | 36.6 ± 0.2 | — | 8.0 | |
| P126D | — | — | — | — | — | — | 21.2 | ||
| Q127D | 0.75 | 4.60 ± 0.46 | 193.8 ± 4.9 | 42.15 ± 4.39 | 36.2 ± 0.4 | — | 10.1 | ||
| Q127G | 0.25 | 6.17 ± 0.27 | 71.3 ± 0.9 | 11.55 ± 0.52 | 36.6 ± 0.2 | — | — | ||
| Q127I | 1.74 | 3.43 ± 0.22 | 152.5 ± 2.3 | 44.49 ± 2.96 | 40.4 ± 0.7 | — | 5.4 | ||
| ► | Q127P | 0.98 ± 0.23 | 5.00 ± 0.37 | 263.9 ± 5.5 | 53.02 ± 4.35 | 39.5 ± 0.4 | — | 336.0 | |
| Q127S | 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 |
||
| ► | Q127T | 0.75 ± 0.54 | 5.54 ± 0.49 | 157.7 ± 3.8 | 28.40 ± 2.58 | 38.0 ± 0.6 | — | 7.5 | |
| W128F | 1.60 | 4.77 ± 0.44 | 47.6 ± 1.1 | 9.97 ± 0.96 | 36.8 ± 0.4 | — | 6.3 | ||
| W128R | 0.18 | 3.71 ± 0.99 | 62.4 ± 4.1 | 16.81 ± 4.60 | 41.3 ± 1.5 | — | 10.1 | ||
| E130D | 0.22 | 2.42 ± 0.32 | 136.6 ± 4.3 | 56.50 ± 7.77 | 39.2 ± 0.3 | — | — | ||
| ► | D131E | 0.35 ± 0.12 | 6.32 ± 0.70 | 561.0 ± 15.8 | 91.96 ± 9.26 | 39.1 ± 0.2 | — | 1.8 | |
| E132Q | 0.90 | 5.17 ± 0.46 | 348.9 ± 8.3 | 67.49 ± 6.21 | — | 43.4 ± 0.0 | -1.1 | ||
| T136Y | 3.22 | 1.58 ± 0.16 | 248.5 ± 6.2 | 157.21 ± 15.94 | — | 44.5 ± 0.2 | 6.8 | ||
| Q142E | 4.17 | 6.47 ± 1.12 | 754.0 ± 35.2 | 116.47 ± 20.92 | — | 47.7 ± 1.3 | -0.6 | ||
| ► | Q142K | 1.94 ± 1.74 | 5.04 ± 0.66 | 168.5 ± 6.1 | 44.04 ± 6.77 | — | 45.0 ± 0.2 | -2.4 | |
| K145R | 0.54 | 4.57 ± 1.26 | 60.2 ± 4.3 | 13.17 ± 3.74 | — | — | — | ||
| Y147W | 1.70 | 2.48 ± 0.47 | 280.5 ± 12.4 | 113.00 ± 22.09 | — | 43.8 ± 0.4 | -8.8 | ||
| ► | A148C | 2.42 | 4.02 ± 0.28 | 368.6 ± 6.5 | 91.74 ± 6.52 | — | 44.3 ± 0.1 | 3.0 | |
| A148E | 2.62 | 4.95 ± 0.38 | 295.1 ± 6.0 | 59.61 ± 4.74 | 40.8 ± 0.1 | — | — | ||
| M152I | 2.60 | 9.03 ± 0.70 | 377.2 ± 8.9 | 41.79 ± 3.38 | — | 42.2 ± 0.1 | 88.5 | ||
| ► | D153V | 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 | |
| E157A | 0.27 | 30.92 ± 13.46 | 112.2 ± 20.0 | 3.63 ± 1.71 | — | — | 1.6 | ||
| E157D | * | 3.46 ± 0.76 | 720.9 ± 39.0 | 208.26 ± 47.19 | 38.8 ± 0.3 | 49.5 ± 0.0 | — | ||
| E157F | 0.72 | 6.49 ± 0.56 | 2,334.7 ± 61.1 | 359.48 ± 32.42 | — | — | — | ||
| N160D | 0.72 | 6.32 ± 0.56 | 330.1 ± 8.2 | 52.25 ± 4.80 | — | 47.5 ± 0.0 | -1.0 | ||
| N160H | 2.06 | 2.17 ± 0.46 | 344.9 ± 16.7 | 159.00 ± 34.33 | — | 44.1 ± 0.0 | -0.4 | ||
| N160T | 0.26 | 2.55 ± 0.59 | 19.7 ± 1.1 | 7.73 ± 1.83 | — | — | — | ||
| N160Y | 1.15 | 7.39 ± 0.93 | 1,508.5 ± 51.8 | 204.18 ± 26.62 | 40.8 ± 0.2 | — | -1.6 | ||
| W161N | 2.79 | 4.95 ± 0.18 | 850.8 ± 7.9 | 171.78 ± 6.35 | — | — | — | ||
| N163A | 3.59 | 14.26 ± 1.61 | 402.4 ± 15.5 | 28.22 ± 3.37 | — | 44.1 ± 0.3 | 2.4 | ||
| N163C | 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 |
||
| N163H | 0.99 | 14.81 ± 0.91 | 144.3 ± 3.0 | 9.74 ± 0.63 | — | 41.2 ± 0.0 | 20.6 | ||
| N163S | 3.61 | 6.71 ± 0.56 | 393.5 ± 9.3 | 58.64 ± 5.07 | — | 42.5 ± 0.1 | — | ||
| ► | N163V | 1.77 ± 0.61 | 6.09 ± 1.22 | 560.4 ± 32.8 | 96.39 ± 27.65 | 38.9 ± 118,735.5 | — | 4.2 | |
| N166A | * | 11.95 ± 0.91 | 6.5 ± 0.0 | 0.65 ± 0.00 | 41.2 ± 0.3 | 44.4 ± 0.0 | — | ||
| N166C | * | 5.42 ± 0.32 | 21.6 ± 0.0 | 3.90 ± 0.22 | 38.0 ± 0.3 | 40.5 ± 0.0 | — | ||
| N166D | * | 15.19 ± 1.41 | 10.8 ± 0.0 | 0.65 ± 0.00 | 38.5 ± 0.3 | — | — | ||
| N166E | — | — | — | — | — | — | — | ||
| N166G | 2.38 | 9.68 ± 0.47 | 2.4 ± 0.0 | 0.25 ± 0.01 | — | 41.7 ± 0.0 | 6.0 | ||
| N166H | * | — | — | — | — | 43.8 ± 0.1 | 21.9 | ||
| N166K | — | — | — | — | — | — | — | ||
| N166Q | 0.10 | 7.01 ± 4.09 | 1.1 ± 0.2 | 0.16 ± 0.10 | — | — | — | ||
| N166S | 0.30 | 2.67 ± 0.41 | 1,758.0 ± 62.2 | 658.99 ± 104.13 | 38.1 ± 0.5 | — | — | ||
| ► | N166T | 0.09 ± 0.06 | 23.01 ± 2.67 | 4.8 ± 0.2 | 0.21 ± 0.03 | — | — | 4.5 | |
| E167A | * | 1.01 ± 0.17 | — | — | 46.0 ± 0.1 | 53.9 ± 0.0 | — | ||
| E167G | — | — | — | — | — | — | — | ||
| E167R | — | — | — | — | — | — | — | ||
| P168A | 0.20 | 3.73 ± 0.33 | 554.7 ± 12.5 | 148.77 ± 13.64 | 39.5 ± 0.2 | — | — | ||
| ► | Y169F | 0.32 ± 0.05 | 5.06 ± 1.13 | 7.3 ± 0.5 | 1.45 ± 0.39 | — | — | — | |
| ► | Y169H | 1.31 ± 0.03 | 18.37 ± 1.30 | 564.1 ± 14.2 | 31.86 ± 2.31 | — | 44.3 ± 0.1 | 3.4 | |
| Y169K | 0.34 | 2.24 ± 0.55 | 1.8 ± 0.1 | 0.80 ± 0.20 | — | 48.4 ± 0.5 | 11.4 | ||
| Y169P | — | — | — | — | — | — | — | ||
| Y169S | 1.64 | 6.67 ± 1.04 | 43.3 ± 1.9 | 6.49 ± 1.05 | — | — | 8.2 | ||
| Y169W | 1.90 | 7.58 ± 0.52 | 9.7 ± 0.2 | 1.28 ± 0.09 | — | — | — | ||
| C170A | * | 14.56 ± 1.27 | 391.8 ± 10.8 | 26.84 ± 2.38 | 39.8 ± 0.5 | 51.1 ± 0.0 | — | ||
| C170D | 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 |
||
| C170E | 0.69 | 5.14 ± 0.20 | 183.5 ± 1.9 | 35.68 ± 1.44 | — | — | — | ||
| ► | C170H | 0.36 | 3.73 ± 0.63 | 168.2 ± 6.2 | 45.12 ± 7.79 | — | — | — | |
| ► | C170N | 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 | |
| C170Q | * | 4.92 ± 0.19 | 413.5 ± 4.3 | 84.00 ± 3.46 | 38.9 ± 0.1 | 48.2 ± 0.0 | — | ||
| ► | C170S | 3.71 ± 1.43 | 71.46 ± 9.54 | 17.0 ± 1.3 | 0.18 ± 0.03 | — | 48.0 ± 0.2 | 3.0 | |
| ► | C170T | 0.77 ± 0.57 | 2.06 ± 0.21 | 344.5 ± 8.4 | 83.71 ± 12.22 | 39.1 ± 0.6 | — | 10.9 | |
| A171N | 0.42 | 7.31 ± 0.40 | 157.0 ± 2.4 | 21.46 ± 1.23 | — | — | — | ||
| S172A | 1.35 | 4.33 ± 0.45 | 515.6 ± 14.8 | 119.07 ± 12.78 | — | 44.8 ± 0.5 | -2.2 | ||
| S172D | 0.13 | 68.87 ± 91.89 | 1.6 ± 1.1 | 0.02 ± 0.03 | — | — | — | ||
| S172N | 0.28 | 10.12 ± 5.32 | 1.6 ± 0.2 | 0.16 ± 0.09 | — | — | — | ||
| ► | I173A | 0.40 ± 0.03 | 2.90 ± 0.17 | 463.5 ± 6.8 | 152.12 ± 9.15 | 37.4 ± 0.1 | — | 4.8 | |
| I173Y | 0.86 | 4.82 ± 0.46 | 432.8 ± 11.0 | 89.75 ± 8.93 | 35.0 ± 0.2 | — | 6.7 | ||
| L174A | * | 11.09 ± 0.42 | 662.4 ± 6.5 | 59.75 ± 2.38 | 38.8 ± 0.5 | 42.9 ± 0.0 | — | ||
| ► | L174F | 5.30 ± 2.22 | 3.41 ± 0.46 | 229.3 ± 5.0 | 83.41 ± 8.37 | — | 50.0 ± 0.0 | 1.6 | |
| L174H | 3.26 | 7.23 ± 0.81 | 256.6 ± 8.3 | 35.48 ± 4.14 | — | 46.7 ± 0.1 | — | ||
| ► | L174I | 0.96 ± 0.40 | 10.55 ± 2.11 | 323.1 ± 17.3 | 35.33 ± 3.27 | — | 43.9 ± 0.1 | 39.3 | |
| L174M | 0.79 | 3.42 ± 0.21 | 247.9 ± 3.7 | 72.51 ± 4.49 | — | 45.5 ± 0.1 | — | ||
| ► | L174Q | 1.20 ± 0.07 | 5.37 ± 0.52 | 315.9 ± 12.3 | 105.54 ± 18.25 | — | 43.0 ± 0.1 | 3.9 | |
| L174R | * | 3.36 ± 0.23 | 331.2 ± 6.5 | 98.50 ± 6.93 | 38.8 ± 0.3 | 44.6 ± 0.0 | — | ||
| ► | L174Y | 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 | |
| ► | Y176L | 0.25 ± 0.22 | 2.09 ± 0.15 | 145.4 ± 5.3 | 92.27 ± 15.72 | — | — | 5.5 | |
| T178A | 0.38 | 10.60 ± 0.79 | 425.0 ± 9.7 | 40.10 ± 3.13 | — | — | — | ||
| T178C | 0.33 | 14.14 ± 1.21 | 2,190.6 ± 61.4 | 154.88 ± 13.95 | — | — | — | ||
| ► | T178H | 0.07 ± 0.03 | 2.55 ± 0.67 | 199.6 ± 13.2 | 127.56 ± 178.06 | 37.6 ± 1.0 | — | 10.0 | |
| T178K | 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 |
||
| T178M | 2.50 | 5.04 ± 0.72 | 283.7 ± 12.1 | 56.30 ± 8.37 | — | 43.1 ± 0.1 | 0.3 | ||
| T178R | * | 3.59 ± 0.15 | 655.9 ± 6.5 | 182.71 ± 7.79 | 38.1 ± 0.1 | — | — | ||
| ► | E180A | 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 | |
| E180C | 1.52 | 6.13 ± 0.26 | 70.8 ± 0.8 | 11.55 ± 0.51 | — | — | — | ||
| ► | E180D | 1.95 ± 0.02 | 5.04 ± 0.58 | 404.1 ± 12.5 | 80.17 ± 9.60 | 37.9 ± 78,411.8 | — | — | |
| E180I | — | — | — | — | — | — | — | ||
| E180K | * | 6.19 ± 0.30 | 454.6 ± 6.5 | 73.39 ± 3.68 | 36.7 ± 0.3 | — | — | ||
| E180L | * | 7.48 ± 0.36 | 549.9 ± 8.7 | 73.60 ± 3.68 | 39.6 ± 0.4 | — | — | ||
| E180Q | 0.58 | 8.17 ± 0.46 | 799.4 ± 13.0 | 97.81 ± 5.78 | — | — | — | ||
| ► | E180T | 2.41 ± 0.31 | 7.59 ± 0.87 | 382.0 ± 11.1 | 49.04 ± 5.71 | — | 43.6 ± 0.1 | — | |
| ► | E180W | 0.29 ± 0.09 | 10.32 ± 1.33 | 61.0 ± 1.5 | 9.64 ± 0.92 | 39.6 ± 0.3 | — | 5.2 | |
| H181A | * | 7.67 ± 0.73 | 93.1 ± 2.2 | 12.12 ± 1.08 | 39.6 ± 0.4 | — | — | ||
| ► | H181C | 0.10 ± 0.06 | 3.90 ± 0.31 | 93.3 ± 4.0 | 45.53 ± 17.40 | — | — | 3.4 | |
| ► | H181F | 0.28 ± 0.16 | 4.72 ± 0.34 | 171.2 ± 4.6 | 46.35 ± 5.58 | — | 40.5 ± 0.1 | -3.4 | |
| H181K | 0.67 | 25.15 ± 1.69 | 22.5 ± 0.6 | 0.90 ± 0.06 | — | — | 8.9 | ||
| ► | H181N | 1.16 ± 1.06 | 4.80 ± 0.68 | 340.4 ± 14.2 | 107.79 ± 42.16 | 39.4 ± 0.2 | — | 3.3 | |
| H181Q | 0.17 | 8.73 ± 0.50 | 231.4 ± 4.0 | 26.51 ± 1.59 | — | — | 2.6 | ||
| ► | H181R | 0.09 | 4.96 ± 0.48 | 48.9 ± 1.4 | 9.85 ± 1.00 | — | — | — | |
| H181S | 0.09 | 14.86 ± 0.61 | 51.0 ± 0.7 | 3.43 ± 0.15 | — | — | — | ||
| ► | H181W | 1.02 ± 0.30 | 7.00 ± 1.17 | 331.7 ± 18.5 | 89.44 ± 20.44 | — | — | 160.8 | |
| H181Y | 0.16 | 4.73 ± 0.24 | 4.0 ± 0.1 | 0.84 ± 0.04 | — | — | -1.2 | ||
| H185K | 5.12 | 4.43 ± 0.35 | 377.2 ± 7.7 | 85.12 ± 6.87 | — | 50.0 ± 0.0 | -0.8 | ||
| H185Q | 5.63 | 5.81 ± 0.38 | 412.3 ± 8.1 | 71.02 ± 4.90 | — | 49.5 ± 0.7 | 2.0 | ||
| H185R | 0.68 | 10.04 ± 0.98 | 720.0 ± 21.1 | 71.68 ± 7.28 | — | — | — | ||
| H185S | 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 |
||
| H185T | 4.02 | 3.72 ± 0.29 | 191.0 ± 9.4 | 51.36 ± 4.74 | — | 49.3 ± 1.2 | 8.1 | ||
| H185V | — | — | — | — | — | — | 6.6 | ||
| H185Y | 2.90 | 8.33 ± 2.15 | 528.0 ± 53.4 | 63.39 ± 17.61 | — | 47.2 ± 0.3 | 3.0 | ||
| E186D | 0.00 | 1.09 ± 0.12 | 885.3 ± 21.5 | 813.55 ± 90.21 | 39.6 ± 0.5 | — | 1.8 | ||
| E186H | 0.86 | 6.40 ± 0.55 | 355.9 ± 8.3 | 55.64 ± 4.93 | — | — | — | ||
| E186T | 4.10 | 4.50 ± 0.25 | 581.3 ± 8.6 | 129.17 ± 7.51 | — | — | — | ||
| N187D | 1.31 | 3.84 ± 0.44 | 1,109.4 ± 32.1 | 288.79 ± 33.88 | 40.4 ± 0.3 | — | -0.2 | ||
| E190A | 8.07 | 5.94 ± 0.37 | 384.4 ± 8.6 | 64.76 ± 4.25 | — | 51.2 ± 0.6 | 2.3 | ||
| E190F | 6.13 | 5.32 ± 0.22 | 40.9 ± 0.5 | 7.69 ± 0.34 | — | 50.8 ± 0.6 | 1.8 | ||
| E190T | 2.84 | 6.15 ± 1.07 | 404.9 ± 20.1 | 65.81 ± 11.85 | — | 49.2 ± 0.8 | 5.4 | ||
| A195S | * | 5.09 ± 0.18 | 775.0 ± 8.7 | 152.19 ± 5.63 | 39.2 ± 0.3 | — | — | ||
| H197G | 0.20 | 1.59 ± 0.30 | 91.5 ± 4.4 | 57.56 ± 11.06 | — | — | 7.0 | ||
| ► | H197L | 0.24 ± 0.05 | 2.77 ± 0.38 | 133.3 ± 5.3 | 58.22 ± 11.21 | 37.4 ± 0.2 | — | 28.9 | |
| ► | H197N | 1.07 ± 0.22 | 4.51 ± 0.43 | 168.8 ± 4.2 | 37.42 ± 3.70 | 38.9 ± 0.4 | — | — | |
| H197Q | 0.16 | 6.08 ± 0.86 | 78.8 ± 3.1 | 12.96 ± 1.90 | 37.2 ± 0.3 | — | — | ||
| H197V | 1.04 | 4.27 ± 0.53 | 194.4 ± 6.3 | 45.52 ± 5.88 | 39.9 ± 0.5 | — | 22.1 | ||
| ► | H197Y | 0.75 ± 0.01 | 5.62 ± 0.80 | 186.8 ± 6.7 | 33.86 ± 4.11 | 38.9 ± 0.3 | — | — | |
| C201A | 3.03 | 4.73 ± 0.47 | 82.9 ± 2.2 | 17.52 ± 1.79 | 39.3 ± 0.2 | — | -0.9 | ||
| C201D | — | — | — | — | — | — | 6.8 | ||
| C201F | — | — | — | — | — | — | 287.9 | ||
| C201G | 0.93 | 1.59 ± 0.34 | 18.0 ± 0.8 | 11.33 ± 2.49 | 37.2 ± 0.4 | — | 4.1 | ||
| C201K | 1.76 | 4.96 ± 0.18 | 84.9 ± 0.8 | 17.13 ± 0.65 | 36.7 ± 1.3 | — | 55.6 | ||
| C201L | 0.68 | — | — | — | — | — | 33.2 | ||
| C201M | 1.24 | 4.93 ± 0.67 | 80.9 ± 2.9 | 16.42 ± 2.31 | 36.1 ± 0.4 | — | 42.9 | ||
| C201N | — | — | — | — | — | — | 12.0 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| C201Q | 0.65 | 1.57 ± 0.32 | 2.8 ± 0.1 | 1.78 ± 0.38 | 40.9 ± 1.1 | — | 16.7 | ||
| C201S | 1.98 | 2.75 ± 0.39 | 113.1 ± 3.8 | 41.19 ± 5.96 | 38.6 ± 0.3 | — | 2.0 | ||
| C201T | 5.49 | 2.27 ± 0.24 | 132.1 ± 3.2 | 58.08 ± 6.26 | 35.9 ± 0.3 | — | 3.2 | ||
| S206V | 0.98 | 3.05 ± 0.46 | 1,281.5 ± 47.5 | 419.79 ± 65.64 | 44.3 ± 0.3 | — | -4.1 | ||
| ► | T215E | 1.21 ± 0.07 | 4.93 ± 0.62 | 384.1 ± 13.2 | 77.91 ± 10.34 | — | 46.5 ± 0.2 | -0.2 | |
| I220S | 1.08 | 5.46 ± 0.66 | 1,280.4 ± 42.3 | 234.72 ± 29.55 | 42.4 ± 0.1 | — | 6.4 | ||
| T221A | * | 6.51 ± 0.94 | 381.0 ± 15.2 | 58.45 ± 8.66 | 40.1 ± 0.1 | 47.5 ± 0.0 | — | ||
| T221G | 1.78 | 6.66 ± 1.34 | 142.0 ± 8.0 | 21.33 ± 4.45 | — | 43.0 ± 0.5 | 8.3 | ||
| ► | T221Q | 2.72 ± 1.68 | 5.49 ± 0.46 | 376.9 ± 8.6 | 34.33 ± 4.21 | — | 46.3 ± 0.7 | 22.8 | |
| ► | T221S | 2.06 ± 0.95 | 22.52 ± 2.93 | 352.7 ± 18.2 | 18.48 ± 3.21 | 41.0 ± 0.2 | 47.5 ± 0.3 | — | |
| L222M | 0.82 | 5.58 ± 0.63 | 248.5 ± 7.6 | 44.54 ± 5.18 | — | 44.5 ± 0.3 | — | ||
| ► | L222Q | 1.02 ± 0.52 | 6.36 ± 2.47 | 719.5 ± 21.5 | 104.07 ± 14.53 | — | — | 6.6 | |
| L222R | 3.02 | 2.79 ± 0.26 | 278.9 ± 6.2 | 100.01 ± 9.44 | 36.6 ± 0.3 | — | — | ||
| L222V | 1.38 | 3.46 ± 0.19 | 225.4 ± 3.1 | 65.05 ± 3.75 | 37.2 ± 0.1 | — | — | ||
| ► | N223A | 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 | |
| ► | N223C | 1.07 ± 0.25 | 4.15 ± 0.51 | 224.1 ± 13.1 | 90.44 ± 33.48 | 36.6 ± 0.3 | — | — | |
| ► | N223D | 1.35 ± 0.19 | 3.21 ± 0.34 | 54.6 ± 3.1 | 34.75 ± 9.98 | 35.3 ± 0.2 | — | — | |
| ► | N223E | 2.36 ± 2.02 | 51.99 ± 9.51 | 202.2 ± 26.7 | 3.06 ± 0.73 | 36.3 ± 0.2 | — | 5.6 | |
| ► | N223F | 1.85 ± 0.73 | 1.17 ± 0.20 | 397.9 ± 9.8 | 546.89 ± 60.45 | 37.9 ± 14,182.7 | — | -2.0 | |
| ► | N223G | 10.31 | 31.79 ± 2.80 | 92.5 ± 3.1 | 2.83 ± 0.31 | 38.9 ± 0.3 | 47.4 ± 0.0 | 5.2 | |
| ► | N223H | 7.46 | 4.81 ± 0.23 | 58.8 ± 0.2 | 11.61 ± 0.63 | 39.1 ± 0.3 | 46.8 ± 0.0 | 0.0 | |
| N223I | * | — | — | — | 36.7 ± 0.1 | — | — | ||
| ► | N223K | 3.11 ± 0.45 | 2.21 ± 0.41 | 351.6 ± 15.0 | 159.19 ± 30.49 | 40.0 ± 0.4 | — | — | |
| ► | N223L | 3.30 ± 1.72 | 10.51 ± 2.96 | 368.6 ± 20.7 | 65.72 ± 10.00 | 37.2 ± 1.0 | — | — | |
| ► | N223M | 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 |
||
| N223P | 3.79 | 9.45 ± 0.46 | 77.1 ± 1.1 | 8.15 ± 0.41 | 35.6 ± 0.2 | — | 53.3 | ||
| ► | N223Q | 3.34 ± 3.05 | 5.28 ± 0.97 | 83.1 ± 6.5 | 17.36 ± 5.35 | 36.4 ± 0.2 | — | 2.6 | |
| ► | N223R | 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 | |
| ► | N223S | 1.18 ± 0.50 | 32.82 ± 2.17 | 68.9 ± 1.8 | 3.27 ± 0.14 | 40.4 ± 0.2 | 47.2 ± 0.1 | — | |
| ► | N223T | 4.29 | 11.90 ± 1.23 | 0.4 ± 0.0 | 0.04 ± 0.00 | 39.7 ± 0.2 | — | — | |
| ► | N223V | 1.30 | 2.98 ± 0.42 | 112.1 ± 3.8 | 37.68 ± 5.46 | 36.6 ± 0.3 | — | 54.0 | |
| ► | N223W | 0.69 ± 0.11 | 0.84 ± 0.22 | 165.3 ± 11.3 | 169.64 ± 46.01 | — | — | — | |
| ► | N223Y | 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 | |
| M224A | * | 6.25 ± 0.60 | 448.1 ± 13.0 | 71.66 ± 7.14 | 40.4 ± 0.3 | 47.2 ± 0.0 | — | ||
| ► | M224K | 0.20 ± 0.03 | 9.48 ± 0.59 | 114.2 ± 2.2 | 12.30 ± 0.88 | — | 46.5 ± 0.1 | 5.5 | |
| M224N | 1.50 | 5.59 ± 0.33 | 246.3 ± 3.9 | 44.07 ± 2.67 | 36.6 ± 0.1 | — | — | ||
| M224Q | 2.84 | 8.03 ± 0.20 | 1,008.5 ± 7.3 | 125.56 ± 3.32 | — | — | — | ||
| M224R | 0.24 | 14.98 ± 0.66 | 25.1 ± 0.4 | 1.67 ± 0.08 | — | — | 3.2 | ||
| E225A | * | 0.63 ± 0.15 | 73.6 ± 4.3 | 116.90 ± 28.58 | 36.9 ± 0.1 | 36.7 ± 0.0 | — | ||
| E225H | * | 8.54 ± 0.53 | 132.1 ± 2.2 | 15.37 ± 1.08 | 34.9 ± 0.2 | 40.2 ± 0.0 | — | ||
| E225K | * | 7.22 ± 0.75 | 88.8 ± 2.2 | 12.34 ± 1.30 | 39.0 ± 0.4 | — | — | ||
| E225Q | * | 12.16 ± 0.65 | 547.7 ± 8.7 | 45.03 ± 2.60 | 39.8 ± 0.3 | — | — | ||
| E225R | * | 2.48 ± 0.44 | 34.6 ± 2.2 | 14.07 ± 2.60 | 39.1 ± 0.2 | — | — | ||
| E225Y | * | 18.43 ± 3.14 | 10.8 ± 0.0 | 0.65 ± 0.22 | 37.1 ± 0.5 | — | — | ||
| ► | H226A | 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 | |
| H226S | 4.60 | 6.49 ± 0.92 | 248.1 ± 9.9 | 38.21 ± 5.61 | — | 40.0 ± 0.1 | 1.2 | ||
| P234K | 2.98 | 4.57 ± 0.42 | 349.8 ± 8.5 | 76.53 ± 7.33 | — | 44.8 ± 0.2 | 0.2 | ||
| D236I | 0.58 | 3.94 ± 0.38 | 33.7 ± 1.1 | 8.55 ± 0.88 | — | — | 16.2 | ||
| A239C | 2.40 | 3.89 ± 0.44 | 202.5 ± 5.9 | 52.09 ± 6.13 | — | 39.4 ± 0.9 | 40.3 | ||
| ► | A239D | — | — | — | — | — | — | 43.0 | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | A239E | * | — | — | — | — | 51.8 ± 0.4 | 160.3 | |
| A239S | 0.13 | — | — | 0.01 ± 0.00 | — | 45.4 ± 0.8 | 6.7 | ||
| ► | A239V | 1.03 ± 0.52 | 0.45 ± 0.31 | 1.2 ± 0.2 | 2.51 ± 1.90 | 36.2 ± 1.3 | — | — | |
| R242C | 1.55 | 6.52 ± 0.41 | 161.3 ± 2.8 | 24.76 ± 1.61 | — | — | 6.7 | ||
| ► | R243A | 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 | |
| R243D | * | 10.82 ± 0.47 | 231.6 ± 4.3 | 21.43 ± 1.08 | 37.2 ± 0.0 | 45.7 ± 0.0 | — | ||
| R243E | — | — | — | — | — | — | — | ||
| ► | R243G | 0.25 ± 0.05 | 23.66 ± 2.61 | 456.7 ± 19.1 | 19.31 ± 2.28 | — | — | — | |
| ► | R243K | 2.20 | 12.92 ± 2.43 | 726.3 ± 37.5 | 64.63 ± 9.16 | 39.5 ± 0.2 | 44.0 ± 0.0 | 2.2 | |
| D244H | 0.28 | 9.97 ± 0.78 | 100.2 ± 2.4 | 10.05 ± 0.82 | — | — | — | ||
| D244R | 0.13 | 2.42 ± 0.46 | 5.1 ± 0.2 | 2.10 ± 0.41 | — | — | 1.3 | ||
| ► | F246H | 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 | |
| I247A | 0.20 | 8.50 ± 1.59 | 451.3 ± 16.6 | 53.12 ± 10.12 | 40.4 ± 0.2 | — | 5.3 | ||
| I247E | * | 5.97 ± 1.04 | 407.0 ± 19.5 | 68.19 ± 12.34 | 37.8 ± 0.1 | 45.8 ± 0.0 | — | ||
| I247K | 3.48 | 14.07 ± 0.87 | 134.4 ± 2.8 | 9.55 ± 0.63 | — | — | 4.3 | ||
| ► | I247M | 2.97 | 2.72 ± 0.59 | 236.0 ± 15.5 | 170.89 ± 111.12 | — | — | 6.8 | |
| I247N | * | 2.15 ± 0.13 | 223.0 ± 4.3 | 103.69 ± 6.49 | 36.8 ± 0.6 | 45.0 ± 0.0 | — | ||
| I247Q | 2.18 | 14.73 ± 0.94 | 1,166.6 ± 24.6 | 79.20 ± 5.31 | — | — | — | ||
| I247T | 3.11 | 4.22 ± 0.17 | 681.8 ± 6.9 | 161.68 ± 6.69 | — | — | — | ||
| I247V | 1.54 | 2.04 ± 0.16 | 518.2 ± 10.0 | 254.43 ± 21.17 | — | — | — | ||
| ► | R249K | 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 | |
| W250V | 0.16 | 7.62 ± 2.81 | 4.2 ± 0.4 | 0.55 ± 0.21 | — | — | — | ||
| F251N | 0.30 | 8.99 ± 2.33 | 8.1 ± 0.6 | 0.91 ± 0.25 | — | 50.3 ± 0.7 | 6.2 | ||
| A252E | — | — | — | — | — | — | — | ||
| ► | E253D | 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 |
||
| F256H | 0.28 | 64.81 ± 42.05 | 297.9 ± 110.9 | 4.60 ± 3.44 | — | — | 3.6 | ||
| ► | F256I | 2.56 ± 1.15 | 12.23 ± 1.70 | 315.8 ± 9.7 | 48.72 ± 5.86 | — | 45.3 ± 0.2 | 5.9 | |
| ► | F256M | 2.17 ± 1.17 | 5.99 ± 0.39 | 448.5 ± 8.2 | 74.93 ± 5.10 | — | 44.7 ± 0.1 | 6.8 | |
| ► | F256Y | 0.69 ± 0.46 | 6.86 ± 0.56 | 275.6 ± 6.6 | 40.53 ± 3.70 | — | 40.9 ± 0.1 | 1.4 | |
| D263E | 0.88 | 10.56 ± 0.84 | 1,349.4 ± 33.7 | 127.83 ± 10.64 | — | — | — | ||
| M264D | — | — | — | — | — | — | — | ||
| W267H | 0.27 | 10.37 ± 1.48 | 124.0 ± 6.0 | 11.96 ± 1.80 | — | 45.2 ± 0.0 | 0.0 | ||
| W267Y | 0.16 | 4.66 ± 2.54 | 24.4 ± 3.5 | 5.23 ± 2.96 | — | — | — | ||
| Y268F | 0.30 | 11.19 ± 3.84 | 5.5 ± 0.6 | 0.49 ± 0.18 | 34.5 ± 0.8 | 30.7 ± 18.7 | -0.6 | ||
| T270E | 2.77 | 4.58 ± 0.47 | 422.5 ± 11.4 | 92.19 ± 9.79 | — | 47.6 ± 0.2 | -1.3 | ||
| Y271K | 1.19 | 3.03 ± 0.36 | 1,237.6 ± 35.5 | 408.00 ± 49.49 | 39.7 ± 0.1 | — | 9.2 | ||
| L272I | 0.21 | 3.77 ± 0.53 | 176.9 ± 6.2 | 46.90 ± 6.80 | 40.4 ± 0.6 | — | 0.4 | ||
| ► | L272V | 0.53 ± 0.10 | 4.17 ± 0.73 | 898.7 ± 40.4 | 215.66 ± 38.85 | 40.0 ± 1.0 | — | 21.6 | |
| ► | N273G | 1.14 ± 0.16 | 7.15 ± 1.88 | 1,307.1 ± 97.1 | 246.14 ± 33.92 | 40.6 ± 0.2 | — | 3.2 | |
| ► | N273W | 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 | |
| ► | G274D | 0.46 ± 0.15 | 1.41 ± 0.22 | 592.0 ± 21.7 | 376.28 ± 63.75 | 37.4 ± 0.2 | — | 4.7 | |
| L275F | 1.23 | 0.80 ± 0.78 | 2.4 ± 0.5 | 3.01 ± 2.99 | — | — | 80.9 | ||
| F277N | 0.70 | 1.96 ± 0.22 | 156.5 ± 4.0 | 79.66 ± 9.10 | 34.9 ± 0.1 | — | — | ||
| Q279K | 0.74 | 3.48 ± 0.47 | 424.1 ± 14.2 | 121.94 ± 16.83 | 34.2 ± 0.9 | — | 4.6 | ||
| G281V | 3.54 | 5.16 ± 1.58 | 463.0 ± 36.2 | 89.76 ± 28.41 | — | — | — | ||
| M283I | 0.54 | 5.70 ± 0.43 | 451.7 ± 9.1 | 79.19 ± 6.22 | — | — | — | ||
| ► | D291E | 2.35 ± 0.06 | 4.26 ± 0.43 | 520.3 ± 13.8 | 114.33 ± 11.87 | — | 44.0 ± 0.1 | 3.6 | |
| F292A | — | — | — | — | — | — | 7.0 | ||
| L293V | 0.39 | 3.65 ± 0.41 | 1,461.3 ± 41.3 | 400.17 ± 46.29 | — | 44.3 ± 0.2 | — | ||
| ► | N296A | 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 |
||
| N296C | * | 19.80 ± 2.08 | 8.7 ± 0.0 | 0.43 ± 0.00 | 38.9 ± 0.4 | 47.0 ± 0.0 | — | ||
| N296D | — | — | — | — | — | — | — | ||
| N296E | * | — | — | — | — | — | 18.9 | ||
| N296F | — | — | — | — | — | — | 86.3 | ||
| ► | N296G | 3.88 ± 2.24 | 24.01 ± 2.80 | 1.1 ± 0.0 | 0.04 ± 0.01 | — | — | — | |
| N296H | * | — | — | — | — | 42.1 ± 0.0 | 162.0 | ||
| N296I | 0.64 | 1.43 ± 0.28 | 1.3 ± 0.1 | 0.92 ± 0.19 | 39.6 ± 1.3 | — | 77.0 | ||
| N296K | — | — | — | — | — | — | — | ||
| N296L | — | — | — | — | — | — | 67.3 | ||
| N296M | * | — | — | — | — | — | 41.9 | ||
| N296P | — | — | — | — | — | — | — | ||
| N296Q | * | — | — | — | — | — | — | ||
| N296R | 0.37 | 0.70 ± 0.15 | 7.9 ± 0.3 | 11.25 ± 2.44 | — | — | — | ||
| ► | N296S | 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 | |
| N296T | 2.11 | 13.62 ± 1.65 | 29.2 ± 1.2 | 2.15 ± 0.27 | 38.3 ± 0.4 | 38.3 ± 0.4 | 11.8 | ||
| ► | N296V | 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 | |
| ► | N296W | — | — | — | — | — | — | 119.6 | |
| ► | N296Y | 0.61 | 48.85 ± 36.83 | 0.6 ± 0.2 | 0.01 ± 0.01 | — | 49.0 ± 0.8 | 87.2 | |
| Y297A | * | 4.98 ± 0.17 | 136.4 ± 2.2 | 27.49 ± 1.08 | 40.4 ± 0.2 | 43.3 ± 0.0 | — | ||
| Y297F | * | 5.99 ± 0.32 | 601.8 ± 8.7 | 100.45 ± 5.63 | 40.8 ± 0.1 | 44.0 ± 0.0 | — | ||
| Y297H | * | — | — | — | — | — | 3.2 | ||
| Y297L | — | — | — | — | — | — | — | ||
| Y298A | * | — | — | — | — | — | — | ||
| Y298G | * | — | — | — | — | — | — | ||
| Y298N | 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 |
||
| ► | T299A | 2.99 | 47.09 ± 4.63 | 0.6 ± 0.0 | 0.01 ± 0.00 | — | — | — | |
| ► | T299C | 0.60 ± 0.43 | 10.65 ± 1.48 | 248.3 ± 15.2 | 21.43 ± 4.20 | — | — | — | |
| T299E | — | — | — | — | — | — | — | ||
| T299F | 5.25 | 0.84 ± 0.14 | 226.7 ± 7.7 | 269.30 ± 46.46 | — | — | 0.1 | ||
| T299H | 0.46 | 8.23 ± 1.10 | 101.5 ± 3.9 | 12.33 ± 1.72 | — | — | — | ||
| T299R | 0.91 | 1.27 ± 0.15 | 780.7 ± 23.1 | 615.34 ± 75.45 | 42.4 ± 0.1 | — | 0.8 | ||
| T299S | 2.46 | 3.91 ± 0.83 | 297.1 ± 18.2 | 76.00 ± 16.89 | — | 43.8 ± 0.3 | 0.4 | ||
| T299W | 2.62 | 0.66 ± 0.10 | 3.0 ± 0.1 | 4.49 ± 0.73 | — | — | 3.0 | ||
| T299Y | 4.35 | 5.95 ± 0.52 | 497.2 ± 12.0 | 83.60 ± 7.60 | — | 45.4 ± 0.0 | -1.1 | ||
| R300A | 0.62 | 2.36 ± 0.49 | 52.2 ± 2.6 | 22.07 ± 4.74 | 41.8 ± 0.1 | — | 5.9 | ||
| S301E | * | 5.28 ± 0.05 | 662.4 ± 2.2 | 125.56 ± 1.30 | 39.6 ± 0.2 | — | — | ||
| S301R | 0.76 | 2.56 ± 1.14 | 435.2 ± 45.8 | 170.03 ± 77.66 | 39.4 ± 0.2 | — | — | ||
| S301T | 0.85 | 3.34 ± 0.37 | 766.1 ± 21.2 | 229.47 ± 26.45 | 39.4 ± 0.3 | — | — | ||
| ► | I302R | 4.56 | 3.48 ± 0.55 | 263.8 ± 10.5 | 75.84 ± 12.43 | — | — | 3.2 | |
| I303N | * | 4.48 ± 0.32 | 569.3 ± 10.8 | 127.07 ± 9.31 | 38.6 ± 0.5 | — | — | ||
| ► | S305A | 2.13 ± 0.03 | 4.83 ± 0.24 | 329.6 ± 4.4 | 68.28 ± 3.52 | — | 44.7 ± 0.0 | -0.8 | |
| S305D | 6.77 | 14.89 ± 1.41 | 74.6 ± 2.4 | 5.01 ± 0.50 | 39.2 ± 0.2 | — | 5.9 | ||
| ► | S305F | 7.84 ± 1.20 | 3.79 ± 0.29 | 124.9 ± 2.6 | 35.00 ± 3.80 | 39.3 ± 0.1 | — | 6.2 | |
| ► | S305H | 3.21 ± 2.02 | 4.81 ± 0.37 | 568.3 ± 14.0 | 133.50 ± 16.70 | — | — | — | |
| S305K | 6.91 | 5.72 ± 0.53 | 197.8 ± 5.0 | 34.56 ± 3.32 | 37.6 ± 0.5 | — | 9.8 | ||
| S305N | 3.27 | 5.43 ± 0.16 | 718.7 ± 5.4 | 132.24 ± 3.92 | — | — | — | ||
| ► | S305W | 5.62 ± 0.69 | 5.41 ± 0.87 | 193.1 ± 12.1 | 38.55 ± 10.43 | 38.8 ± 0.3 | — | 6.2 | |
| S305Y | 11.07 | 5.53 ± 0.48 | 153.3 ± 3.7 | 27.74 ± 2.52 | 39.9 ± 0.1 | — | 5.8 | ||
| T306K | 1.17 | 3.45 ± 0.37 | 865.0 ± 23.3 | 250.97 ± 27.94 | 42.0 ± 0.1 | — | 76.0 | ||
| A309K | 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 |
||
| ► | A309N | 1.68 ± 0.09 | 4.97 ± 0.19 | 340.9 ± 3.5 | 68.54 ± 2.68 | — | 44.6 ± 0.1 | -2.1 | |
| ► | A309Q | 2.08 | 3.75 ± 0.63 | 587.8 ± 25.0 | 156.87 ± 27.23 | 38.3 ± 0.8 | — | — | |
| ► | A309S | 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 | |
| V314D | 3.05 | 6.84 ± 0.65 | 295.0 ± 8.0 | 43.14 ± 4.25 | — | 41.8 ± 0.1 | 2.6 | ||
| ► | E315V | 0.87 ± 0.12 | 3.37 ± 0.18 | 958.4 ± 15.2 | 287.10 ± 19.44 | 38.6 ± 0.2 | — | 3.7 | |
| Q316A | 1.25 | 4.45 ± 0.57 | 603.3 ± 19.7 | 135.49 ± 17.81 | 39.6 ± 0.1 | — | — | ||
| ► | Q316C | 3.14 ± 1.54 | 5.54 ± 0.46 | 285.0 ± 5.3 | 49.25 ± 3.80 | — | — | 0.9 | |
| Q316E | 4.20 | 6.76 ± 0.86 | 413.8 ± 14.9 | 61.19 ± 8.05 | 39.7 ± 0.1 | — | — | ||
| Q316L | 3.90 | 3.58 ± 0.23 | 363.5 ± 5.8 | 101.67 ± 6.76 | 40.1 ± 0.1 | — | — | ||
| Q316N | 0.37 | 6.14 ± 0.89 | 291.1 ± 11.4 | 47.42 ± 7.10 | — | — | — | ||
| Q316P | 2.63 | 3.48 ± 0.55 | 126.7 ± 4.7 | 36.42 ± 5.92 | — | — | 4.3 | ||
| Q316R | * | 3.58 ± 0.51 | 565.0 ± 19.5 | 157.82 ± 23.16 | 42.1 ± 0.3 | — | — | ||
| Q316V | 2.53 | 8.77 ± 0.68 | 900.2 ± 20.5 | 102.63 ± 8.35 | — | — | — | ||
| V317A | 2.85 | 2.64 ± 0.29 | 182.3 ± 4.8 | 69.19 ± 7.93 | 37.9 ± 0.4 | — | 2.8 | ||
| V317D | 8.32 | 5.14 ± 0.61 | 126.2 ± 4.1 | 24.58 ± 3.05 | 38.1 ± 0.2 | — | 6.7 | ||
| V317E | 6.38 | 2.42 ± 0.34 | 125.9 ± 4.4 | 52.01 ± 7.45 | 38.0 ± 0.1 | — | 2.8 | ||
| V317F | 5.00 | 4.47 ± 0.33 | 210.1 ± 4.0 | 46.98 ± 3.54 | 37.8 ± 0.2 | — | 8.2 | ||
| V317H | 6.95 | 3.43 ± 0.33 | 227.5 ± 5.8 | 66.32 ± 6.57 | 38.2 ± 0.1 | — | 5.6 | ||
| V317K | 2.94 | 5.55 ± 0.83 | 269.1 ± 12.3 | 48.45 ± 7.60 | 36.0 ± 0.4 | — | 2.4 | ||
| ► | V317L | 8.02 ± 0.87 | 1.82 ± 0.33 | 151.2 ± 5.2 | 108.02 ± 17.69 | 38.8 ± 0.3 | — | 2.3 | |
| V317M | 7.63 | 6.76 ± 0.43 | 181.4 ± 3.3 | 26.84 ± 1.79 | 38.5 ± 0.2 | — | 4.3 | ||
| V317Q | 6.59 | 3.16 ± 0.49 | 165.6 ± 6.3 | 52.36 ± 8.31 | 38.5 ± 0.2 | — | 2.4 | ||
| V317T | 2.24 | 3.44 ± 0.37 | 164.7 ± 4.5 | 47.91 ± 5.38 | 38.5 ± 0.3 | — | 3.3 | ||
| V317W | 6.07 | 4.76 ± 0.20 | 168.1 ± 1.9 | 35.36 ± 1.58 | 37.1 ± 0.2 | — | 7.7 | ||
| ► | H318C | 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 |
||
| H318D | 0.32 | 0.68 ± 0.24 | 897.0 ± 54.3 | 1,312.84 ± 460.56 | 40.1 ± 0.1 | — | — | ||
| ► | H318E | 1.78 | 2.84 ± 0.39 | 183.7 ± 6.8 | 64.70 ± 9.19 | — | 47.2 ± 0.1 | -1.2 | |
| H318I | 3.82 | 6.73 ± 0.32 | 588.1 ± 7.6 | 87.37 ± 4.25 | — | — | — | ||
| H318K | 0.38 | 6.14 ± 0.56 | 36.6 ± 0.9 | 5.96 ± 0.57 | — | — | — | ||
| ► | H318N | 0.92 ± 0.26 | 1.35 ± 0.25 | 494.2 ± 20.3 | 393.33 ± 86.03 | 40.8 ± 0.3 | — | -1.5 | |
| ► | H318P | 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 | |
| H318Q | 0.41 | 1.12 ± 0.78 | 6.6 ± 1.1 | 5.88 ± 4.26 | 40.4 ± 0.5 | — | -1.1 | ||
| H318T | 4.76 | 4.67 ± 0.11 | 658.5 ± 4.0 | 141.04 ± 3.48 | — | — | — | ||
| H318Y | 0.90 | 7.02 ± 0.66 | 347.3 ± 9.4 | 49.47 ± 4.85 | — | 45.7 ± 0.1 | — | ||
| M319K | 0.35 | 3.77 ± 0.90 | 439.2 ± 25.9 | 116.43 ± 28.63 | — | — | — | ||
| E321R | 3.49 | 8.52 ± 0.97 | 5.3 ± 0.2 | 0.62 ± 0.07 | — | — | — | ||
| T324S | 1.84 | 9.44 ± 2.87 | 422.2 ± 42.1 | 44.72 ± 14.29 | — | 43.3 ± 0.4 | 2.4 | ||
| M326A | * | 9.34 ± 0.80 | 342.0 ± 8.7 | 36.59 ± 3.25 | 37.1 ± 0.1 | 42.5 ± 0.0 | — | ||
| M326C | 0.32 | 16.52 ± 1.78 | 105.5 ± 4.0 | 6.39 ± 0.73 | 39.9 ± 0.3 | — | 4.5 | ||
| M326D | 1.11 | 4.44 ± 0.89 | 5.4 ± 0.3 | 1.21 ± 0.25 | — | 40.3 ± 0.1 | 5.4 | ||
| ► | M326I | 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 | |
| M326K | — | — | — | — | — | — | — | ||
| ► | M326S | 0.17 ± 0.00 | 10.41 ± 1.33 | 503.0 ± 17.2 | 58.42 ± 5.44 | 38.6 ± 0.4 | — | 5.2 | |
| ► | W328A | 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 | |
| W328C | * | 4.18 ± 0.53 | 8.7 ± 0.0 | 2.16 ± 0.22 | 39.3 ± 0.2 | 41.1 ± 0.0 | — | ||
| W328D | 2.22 | 27.09 ± 3.65 | 1,039.5 ± 55.4 | 38.38 ± 5.57 | — | — | — | ||
| W328E | 3.58 | 2.26 ± 0.28 | 1.6 ± 0.1 | 0.71 ± 0.09 | 38.0 ± 0.3 | — | 5.2 | ||
| W328F | 1.87 | 12.63 ± 3.27 | 116.0 ± 9.7 | 9.19 ± 2.50 | — | — | — | ||
| ► | W328G | * | — | — | — | 39.0 ± 0.6 | — | 10.0 | |
| W328H | * | 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 |
||
| W328I | 3.45 | 1.29 ± 0.16 | 6.2 ± 0.2 | 4.84 ± 0.61 | 39.6 ± 0.5 | — | 29.7 | ||
| ► | W328K | 0.74 ± 0.45 | 3.38 ± 0.88 | 5.9 ± 0.4 | 1.61 ± 0.38 | 39.4 ± 0.4 | — | 3.6 | |
| W328L | * | 5.74 ± 0.35 | 88.8 ± 2.2 | 15.37 ± 1.08 | 40.4 ± 0.5 | — | — | ||
| ► | W328M | 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 | |
| W328N | 3.86 | 1.93 ± 0.15 | 4.4 ± 0.1 | 2.27 ± 0.18 | 40.4 ± 0.4 | — | 6.2 | ||
| W328P | — | — | — | — | — | — | 112.0 | ||
| ► | W328Q | 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 | |
| ► | W328R | 2.05 | 14.18 ± 3.12 | 213.4 ± 15.7 | 15.05 ± 3.49 | 40.0 ± 0.2 | 42.7 ± 0.0 | 3.0 | |
| W328S | 1.89 | 1.05 ± 0.21 | 6.1 ± 0.2 | 5.81 ± 1.17 | 42.0 ± 1.5 | — | 7.2 | ||
| W328T | 3.96 | 2.03 ± 0.26 | 7.0 ± 0.2 | 3.45 ± 0.46 | 38.2 ± 0.5 | — | 8.9 | ||
| ► | W328V | 2.59 ± 0.91 | 1.43 ± 0.16 | 8.6 ± 0.2 | 6.04 ± 0.71 | 39.1 ± 0.7 | — | 41.5 | |
| ► | W328Y | 3.02 | 2.41 ± 0.20 | 32.6 ± 0.6 | 13.52 ± 1.16 | 40.8 ± 0.2 | — | 5.1 | |
| H331A | 0.77 | 8.24 ± 0.32 | 423.2 ± 4.9 | 51.36 ± 2.09 | — | 45.4 ± 0.1 | — | ||
| ► | H331K | 1.83 ± 0.26 | 6.18 ± 2.38 | 306.7 ± 35.1 | 81.33 ± 27.49 | 39.0 ± 0.3 | — | 1.1 | |
| H331P | 0.36 | 2.71 ± 0.67 | 4.3 ± 0.2 | 1.58 ± 0.40 | — | — | 51.0 | ||
| P332N | — | — | — | — | — | — | — | ||
| S334A | * | 4.34 ± 0.11 | 668.9 ± 4.3 | 154.13 ± 4.11 | 39.3 ± 0.3 | 41.0 ± 0.0 | — | ||
| S334G | 2.43 | 5.82 ± 0.16 | 1,425.9 ± 14.4 | 244.86 ± 7.02 | — | — | — | ||
| S334T | 0.89 | 4.50 ± 1.38 | 27.4 ± 1.9 | 6.09 ± 1.91 | — | — | — | ||
| ► | Y336F | 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 | |
| K337D | 1.20 | 6.11 ± 0.90 | 279.9 ± 11.1 | 45.78 ± 6.95 | — | — | — | ||
| L339S | 0.28 | 8.02 ± 0.47 | 189.2 ± 3.1 | 23.58 ± 1.43 | — | — | — | ||
| ► | E343A | 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 | |
| E343S | 0.81 | 3.54 ± 0.46 | 308.7 ± 10.1 | 87.21 ± 11.72 | — | 45.0 ± 0.3 | 4.4 | ||
| ► | K344A | 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 |
||
| ► | F346E | 0.10 | 45.41 ± 19.90 | 29.0 ± 6.2 | 0.64 ± 0.31 | — | — | -3.1 | |
| F346S | — | — | — | — | — | — | — | ||
| ► | L353I | 2.54 ± 0.87 | 4.71 ± 0.43 | 281.6 ± 8.4 | 50.60 ± 5.75 | — | 47.7 ± 0.1 | 0.1 | |
| I354V | 3.95 | 4.78 ± 0.39 | 436.1 ± 9.2 | 91.20 ± 7.74 | — | — | — | ||
| ► | T355A | 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 | |
| T355G | 0.43 | 6.83 ± 0.37 | 21.1 ± 0.4 | 3.09 ± 0.18 | — | — | 4.4 | ||
| T355P | 0.24 | 5.21 ± 0.36 | 252.6 ± 4.6 | 48.50 ± 3.42 | — | — | — | ||
| ► | T355S | 0.23 | 5.11 ± 1.38 | 234.4 ± 18.2 | 45.83 ± 12.86 | 37.5 ± 0.3 | — | — | |
| ► | T355V | 0.05 ± 0.01 | 2.68 ± 0.45 | 79.1 ± 3.1 | 29.52 ± 5.06 | — | — | — | |
| T355W | — | — | — | — | — | — | 71.5 | ||
| E356A | * | — | — | — | — | — | — | ||
| E356D | 0.45 | 2.35 ± 0.99 | 1.6 ± 0.2 | 0.68 ± 0.29 | — | — | — | ||
| N357A | * | 5.38 ± 0.67 | 2.2 ± 0.0 | 0.43 ± 0.00 | 40.8 ± 0.4 | 46.7 ± 0.0 | — | ||
| ► | N357S | 0.63 ± 0.38 | 3.34 ± 0.44 | 54.6 ± 2.2 | 20.46 ± 4.29 | — | 40.9 ± 0.3 | 1.8 | |
| G358A | — | — | — | — | — | — | — | ||
| G358N | 0.31 | 8.65 ± 2.96 | 0.4 ± 0.0 | 0.04 ± 0.01 | — | — | -3.6 | ||
| A359T | 1.90 | 2.89 ± 0.18 | 285.4 ± 4.2 | 98.63 ± 6.22 | — | 45.1 ± 0.1 | — | ||
| M361T | * | 4.83 ± 0.48 | 357.2 ± 8.7 | 74.04 ± 7.58 | 39.5 ± 0.2 | — | — | ||
| L365M | * | 5.20 ± 0.19 | 751.2 ± 6.5 | 144.39 ± 5.41 | 40.0 ± 0.2 | — | — | ||
| V366N | 2.07 | 4.64 ± 0.32 | 478.2 ± 8.3 | 103.16 ± 7.24 | — | — | — | ||
| ► | V366Q | 1.80 ± 0.67 | 5.67 ± 0.41 | 202.7 ± 6.4 | 36.86 ± 4.52 | — | 42.0 ± 0.4 | 1.6 | |
| G368N | 3.39 | 4.77 ± 0.12 | 961.6 ± 8.5 | 201.64 ± 5.33 | — | — | — | ||
| R375F | 0.41 | 4.84 ± 1.70 | 1.0 ± 0.1 | 0.21 ± 0.08 | — | — | 5.8 | ||
| R375H | 3.57 | 3.73 ± 0.33 | 625.2 ± 14.0 | 167.65 ± 15.29 | — | — | 2.9 | ||
| R375W | 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 |
||
| R375Y | 0.42 | 2.88 ± 0.47 | 1.7 ± 0.1 | 0.58 ± 0.10 | — | — | 6.8 | ||
| H376R | * | 6.31 ± 0.30 | 580.2 ± 6.5 | 92.00 ± 4.55 | 39.2 ± 0.3 | 51.4 ± 0.0 | — | ||
| Y378H | 0.49 | 5.45 ± 0.36 | 418.7 ± 7.4 | 76.84 ± 5.30 | — | — | — | ||
| Y378T | 0.43 | — | — | 0.09 ± 0.00 | — | — | 8.2 | ||
| E380Q | 0.31 | 4.86 ± 0.69 | 1,537.4 ± 58.0 | 316.60 ± 46.56 | 39.6 ± 0.2 | — | — | ||
| E380S | 1.11 | 4.64 ± 0.69 | 590.6 ± 22.4 | 127.35 ± 19.44 | — | — | — | ||
| H382R | * | — | — | — | — | — | — | ||
| H382T | — | — | — | — | — | — | — | ||
| ► | H382Y | 0.61 ± 0.25 | 2.52 ± 0.99 | 200.8 ± 18.9 | 628.48 ± 1,425.52 | 39.3 ± 0.3 | — | -0.9 | |
| H387A | 1.09 | 6.22 ± 0.42 | 644.9 ± 11.9 | 103.69 ± 7.30 | — | — | — | ||
| ► | H387I | 3.42 ± 0.47 | 2.73 ± 0.31 | 183.2 ± 5.2 | 68.95 ± 8.16 | — | 40.2 ± 0.2 | 12.9 | |
| H387R | 0.43 | 6.00 ± 0.39 | 31.4 ± 0.5 | 5.24 ± 0.35 | — | — | — | ||
| I390K | 0.17 | 9.29 ± 1.08 | 94.5 ± 3.4 | 10.18 ± 1.24 | — | 43.6 ± 0.4 | 4.7 | ||
| E391F | 3.62 | 4.79 ± 0.37 | 317.7 ± 6.5 | 66.28 ± 5.31 | — | — | — | ||
| E391G | 5.11 | 1.62 ± 0.21 | 177.9 ± 5.0 | 109.58 ± 14.28 | — | 44.6 ± 0.1 | -4.4 | ||
| ► | E391H | 0.77 ± 0.03 | 3.94 ± 0.60 | 245.5 ± 17.2 | 111.21 ± 19.28 | 40.1 ± 0.2 | — | -3.3 | |
| ► | E391L | 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 | |
| E391R | 4.14 | 3.67 ± 0.80 | 304.6 ± 16.4 | 82.96 ± 18.62 | — | — | — | ||
| E392R | * | — | — | — | — | 45.2 ± 0.1 | -0.7 | ||
| Q395N | 0.47 | 4.84 ± 1.10 | 331.4 ± 21.1 | 68.41 ± 16.07 | — | — | — | ||
| Y399C | — | — | — | — | — | — | 0.9 | ||
| Y399D | — | — | — | — | — | — | 9.4 | ||
| Y399E | — | — | — | — | — | — | 11.3 | ||
| Y399F | 1.14 | 6.40 ± 1.03 | 155.0 ± 7.0 | 24.23 ± 4.03 | 38.0 ± 0.2 | — | 1.8 | ||
| Y399G | — | — | — | — | — | — | 14.6 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | Y399P | — | — | — | — | — | — | 283.8 | |
| Y399Q | — | — | — | — | — | — | 9.0 | ||
| Y399R | — | — | — | — | — | — | 9.3 | ||
| Y399S | — | — | — | — | — | — | 8.2 | ||
| Y399V | — | — | — | — | — | — | 21.5 | ||
| Y399W | 1.28 | 1.96 ± 0.54 | 14.1 ± 1.1 | 7.20 ± 2.05 | 38.4 ± 0.5 | — | 3.9 | ||
| F400A | 0.68 | 2.78 ± 0.61 | 25.1 ± 1.4 | 9.03 ± 2.03 | 39.2 ± 4.4 | 39.1 ± 2.9 | 6.7 | ||
| F400D | — | — | — | — | — | — | 7.9 | ||
| F400E | — | — | — | — | — | — | 0.0 | ||
| F400G | — | — | — | — | — | — | 12.0 | ||
| F400H | — | — | — | — | — | — | 3.8 | ||
| F400I | 2.20 | 2.02 ± 0.28 | 79.4 ± 2.5 | 39.24 ± 5.56 | 37.5 ± 0.1 | 37.2 ± 0.8 | 26.9 | ||
| F400K | — | — | — | — | — | — | 0.0 | ||
| F400N | 0.71 | 8.68 ± 0.95 | 86.3 ± 3.1 | 9.94 ± 1.14 | 38.5 ± 0.8 | 38.5 ± 0.8 | 6.0 | ||
| F400R | — | — | — | — | — | — | 8.6 | ||
| F400S | 1.10 | 4.81 ± 0.23 | 88.0 ± 1.1 | 18.31 ± 0.91 | 35.7 ± 0.7 | 35.7 ± 0.7 | 7.6 | ||
| F400V | 0.50 | 3.01 ± 0.44 | 7.1 ± 0.2 | 2.38 ± 0.36 | 36.7 ± 0.3 | — | 27.7 | ||
| F400W | 1.37 | 9.02 ± 0.79 | 105.0 ± 3.3 | 11.64 ± 1.09 | 38.2 ± 0.4 | 38.2 ± 0.4 | 0.5 | ||
| F400Y | — | — | — | — | — | — | 1.6 | ||
| V401N | 0.41 | 4.11 ± 0.25 | 400.5 ± 6.3 | 97.37 ± 6.16 | 34.3 ± 0.2 | — | 4.6 | ||
| W402A | * | — | — | — | — | — | — | ||
| W402C | * | 70.33 ± 5.89 | 2.2 ± 0.0 | — | 41.7 ± 0.1 | 44.8 ± 0.0 | — | ||
| W402G | * | — | — | — | — | — | — | ||
| W402Q | 3.85 | 21.84 ± 3.20 | 0.6 ± 0.0 | 0.03 ± 0.00 | — | 44.8 ± 0.1 | 6.4 | ||
| W402R | * | 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 |
||
| W402S | * | — | — | — | — | — | — | ||
| S403A | * | 3.22 ± 0.22 | 435.1 ± 6.5 | 135.09 ± 9.53 | 39.9 ± 0.3 | 47.3 ± 0.0 | — | ||
| S403G | 2.18 | 30.88 ± 2.17 | 287.8 ± 8.3 | 9.32 ± 0.71 | — | — | — | ||
| S403W | — | — | — | — | — | — | 124.8 | ||
| F404K | 0.63 | — | — | — | — | 44.9 ± 0.8 | 18.4 | ||
| L405C | 5.89 | 3.12 ± 0.21 | 744.3 ± 24.0 | 238.68 ± 17.68 | — | — | — | ||
| L405E | 0.27 | 2.58 ± 0.14 | 43.7 ± 0.6 | 16.96 ± 0.95 | — | — | — | ||
| D406A | — | — | — | — | — | — | — | ||
| ► | N407A | 3.83 | 7.75 ± 1.29 | 2.8 ± 0.1 | 0.32 ± 0.04 | 41.9 ± 0.1 | 54.3 ± 0.0 | — | |
| ► | N407C | 1.55 | 18.77 ± 3.31 | 78.0 ± 4.5 | 4.25 ± 0.70 | 43.2 ± 0.2 | 46.0 ± 0.0 | -1.9 | |
| ► | N407D | 1.44 ± 0.25 | 27.12 ± 2.51 | 284.8 ± 14.9 | 62.38 ± 13.36 | — | 42.4 ± 1.8 | 5.7 | |
| ► | N407E | 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 | |
| ► | N407L | 1.90 ± 0.75 | 30.74 ± 2.08 | 22.8 ± 0.6 | 0.74 ± 0.05 | — | 44.6 ± 0.1 | -1.6 | |
| ► | N407M | 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 | |
| ► | N407Q | 2.24 ± 1.29 | 2.48 ± 0.81 | 0.2 ± 0.0 | 0.14 ± 0.12 | — | 48.3 ± 0.3 | 0.5 | |
| ► | N407S | 2.86 ± 2.10 | 26.79 ± 4.28 | 35.2 ± 1.7 | 1.28 ± 0.20 | 44.5 ± 0.1 | — | — | |
| E409A | * | — | — | — | 40.1 ± 0.7 | 49.0 ± 0.0 | — | ||
| ► | E409D | 0.49 | 29.16 ± 4.17 | 28.4 ± 1.5 | 0.94 ± 0.17 | 40.4 ± 0.2 | 42.2 ± 0.0 | -4.4 | |
| E409F | 0.64 | — | — | — | — | — | — | ||
| E409H | 2.36 | 0.02 ± 0.04 | 0.0 ± 0.0 | 1.80 ± 2.83 | — | — | 4.5 | ||
| E409K | * | — | — | — | — | 44.1 ± 0.3 | 25.2 | ||
| ► | E409Q | 3.81 ± 1.18 | 58.18 ± 5.12 | 36.9 ± 3.0 | 0.37 ± 0.07 | — | 47.2 ± 1.3 | 4.3 | |
| E409T | 3.40 | 2.57 ± 1.38 | 15.6 ± 1.9 | 6.05 ± 3.33 | — | — | — | ||
| ► | W410A | 0.91 | 27.09 ± 4.32 | 0.6 ± 0.0 | 0.02 ± 0.00 | — | 42.4 ± 0.2 | — | |
| ► | W410D | 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 |
||
| W410E | — | — | — | — | — | — | 10.6 | ||
| ► | W410F | 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 | |
| W410G | — | — | — | — | — | — | — | ||
| ► | W410H | 0.97 ± 0.47 | 5.58 ± 3.04 | 0.5 ± 0.1 | 1.02 ± 0.79 | — | — | — | |
| ► | W410K | 0.09 | 17.69 ± 3.06 | 3.6 ± 0.2 | 0.20 ± 0.04 | — | — | 3.5 | |
| W410L | * | — | — | — | — | — | 29.5 | ||
| ► | W410N | 0.20 ± 0.05 | — | — | — | — | — | -0.9 | |
| W410Q | — | — | — | — | — | — | — | ||
| ► | W410R | — | — | — | — | — | — | 8.8 | |
| W410S | — | — | — | — | — | — | -1.8 | ||
| W410Y | — | — | — | — | — | — | — | ||
| A411F | 0.31 | 37.97 ± 8.36 | 341.9 ± 33.2 | 9.00 ± 2.17 | — | — | — | ||
| ► | A411H | 1.16 ± 0.57 | 13.31 ± 0.87 | 258.9 ± 5.6 | 19.45 ± 1.36 | — | 43.7 ± 0.1 | 3.0 | |
| ► | A411K | 1.95 ± 1.11 | 5.34 ± 0.67 | 478.2 ± 16.0 | 89.76 ± 10.92 | — | 46.4 ± 0.2 | — | |
| A411S | 0.23 | 9.32 ± 2.00 | 158.5 ± 10.3 | 17.02 ± 3.81 | 38.4 ± 0.2 | — | 1.9 | ||
| A411T | 0.14 | 12.90 ± 2.77 | 153.9 ± 10.7 | 11.93 ± 2.69 | 37.3 ± 0.5 | — | 4.9 | ||
| ► | W412A | 1.93 ± 1.93 | 7.15 ± 1.40 | 184.0 ± 10.8 | 25.77 ± 4.75 | 40.2 ± 0.3 | — | 4.0 | |
| W412C | 6.00 | 2.44 ± 0.13 | 156.2 ± 1.9 | 64.01 ± 3.39 | 39.4 ± 2.2 | — | 5.0 | ||
| ► | W412D | 1.48 ± 1.51 | 25.96 ± 7.34 | 592.0 ± 31.6 | 182.58 ± 18.18 | 40.3 ± 0.3 | — | 6.8 | |
| ► | W412E | 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 | |
| ► | W412F | 0.62 ± 0.16 | 3.45 ± 0.25 | 605.1 ± 11.0 | 175.69 ± 13.06 | 39.6 ± 0.2 | — | 1.2 | |
| W412G | 0.44 | 3.85 ± 0.82 | 14.1 ± 0.7 | 3.66 ± 0.81 | — | — | — | ||
| ► | W412H | 1.39 ± 1.05 | 2.70 ± 0.26 | 375.1 ± 11.0 | 136.73 ± 18.14 | 38.9 ± 0.6 | — | 2.1 | |
| ► | W412I | 1.13 ± 0.00 | 6.88 ± 1.32 | 375.4 ± 31.9 | 41.51 ± 11.89 | 40.3 ± 0.3 | — | 8.2 | |
| W412K | 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 |
||
| W412L | 6.10 | 2.05 ± 0.16 | 87.5 ± 1.6 | 42.57 ± 3.43 | 39.5 ± 0.3 | — | 9.5 | ||
| W412M | 0.62 | 7.11 ± 2.14 | 35.4 ± 2.6 | 4.98 ± 1.55 | 40.0 ± 0.1 | — | -0.6 | ||
| ► | W412N | 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 | |
| W412P | — | — | — | — | — | — | 82.7 | ||
| ► | W412R | 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 | |
| W412S | 6.17 | 4.52 ± 0.76 | 143.7 ± 6.3 | 31.79 ± 5.51 | 39.4 ± 0.4 | — | 4.1 | ||
| ► | W412T | 0.95 ± 0.83 | 3.68 ± 0.85 | 214.6 ± 14.1 | 90.81 ± 29.37 | 36.4 ± 1.3 | — | -356.9 | |
| W412V | 4.17 | 9.62 ± 1.08 | 139.5 ± 4.8 | 14.50 ± 1.71 | 39.3 ± 0.9 | — | 5.0 | ||
| ► | W412Y | 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 | |
| ► | Y414R | 0.28 ± 0.13 | 1.99 ± 0.63 | 50.3 ± 3.3 | 32.27 ± 11.15 | — | — | — | |
| K416A | 3.15 | 7.32 ± 0.60 | 381.9 ± 9.0 | 52.18 ± 4.42 | — | 48.8 ± 0.1 | — | ||
| K416Q | 3.02 | 5.41 ± 0.62 | 386.6 ± 12.1 | 71.53 ± 8.55 | — | 45.8 ± 1.7 | 5.0 | ||
| K416R | 0.41 | 0.85 ± 0.29 | 470.8 ± 32.0 | 555.04 ± 196.18 | 40.1 ± 0.1 | — | — | ||
| R417K | 0.55 | 26.88 ± 10.87 | 399.9 ± 66.3 | 14.88 ± 6.50 | 40.6 ± 0.6 | — | 1.6 | ||
| F418A | * | 16.63 ± 4.00 | — | — | 40.5 ± 0.3 | 44.7 ± 0.0 | — | ||
| F418H | 0.47 | 0.78 ± 0.34 | 9.2 ± 1.0 | 11.81 ± 5.23 | — | — | 2.3 | ||
| F418N | * | — | — | — | — | — | — | ||
| ► | F418Y | 2.11 ± 0.13 | 9.03 ± 0.99 | 202.3 ± 7.4 | 21.64 ± 2.64 | — | 41.6 ± 0.2 | — | |
| I420H | 1.25 | 2.81 ± 0.87 | 4.2 ± 0.3 | 1.51 ± 0.48 | — | 44.5 ± 0.8 | 8.6 | ||
| ► | H422Y | 3.27 ± 0.04 | 4.88 ± 0.44 | 742.6 ± 14.1 | 143.16 ± 11.99 | — | 49.1 ± 1.1 | -2.1 | |
| ► | N424D | 0.79 ± 0.24 | 3.26 ± 0.33 | 1,113.3 ± 26.0 | 356.21 ± 42.67 | 41.3 ± 0.2 | — | -0.2 | |
| Y425F | 2.83 | 30.66 ± 5.28 | 183.8 ± 13.5 | 6.00 ± 1.12 | — | 47.4 ± 0.2 | -0.4 | ||
| E426S | * | 6.60 ± 0.42 | 530.4 ± 34.6 | 80.31 ± 7.36 | 39.5 ± 0.1 | 44.8 ± 0.0 | — | ||
| T427H | 3.64 | 6.10 ± 0.34 | 39.6 ± 0.6 | 6.50 ± 0.37 | — | 46.7 ± 0.2 | 1.7 | ||
| ► | R430H | 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 |
||
| ► | R430L | 1.00 ± 0.85 | 4.71 ± 0.38 | 377.3 ± 8.0 | 80.49 ± 6.64 | — | 44.7 ± 0.2 | 4.9 | |
| R430W | 0.31 | 0.98 ± 0.27 | 35.6 ± 2.0 | 36.11 ± 10.00 | 40.2 ± 0.1 | — | — | ||
| P432A | 2.51 | 9.81 ± 1.03 | 327.2 ± 10.7 | 33.35 ± 3.65 | — | — | — | ||
| ► | L437S | 1.73 ± 0.76 | 1.69 ± 0.18 | 221.9 ± 9.6 | 244.50 ± 85.67 | — | 43.0 ± 0.0 | 1.2 | |
| ► | W438Y | 0.48 | 7.41 ± 0.72 | 823.9 ± 23.2 | 111.17 ± 11.22 | 40.2 ± 0.2 | — | — | |
| A444V | 2.07 | 4.65 ± 0.10 | 859.5 ± 4.6 | 184.98 ± 4.01 | — | — | — |