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.
439 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.
439 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 | 2.06 ± 1.64 | 6.15 ± 1.45 | 782.9 ± 134.3 | 144.56 ± 50.53 | 39.4 ± 0.7 | 45.4 ± 1.0 | — | |
| W11F | 0.71 | 5.33 ± 0.40 | 836.9 ± 16.4 | 156.97 ± 12.12 | — | — | — | ||
| G12N | — | — | — | — | — | — | — | ||
| T13K | * | — | — | — | — | — | 36.0 | ||
| ► | T13S | 0.71 ± 0.51 | 11.02 ± 1.10 | 809.2 ± 36.0 | 153.00 ± 19.79 | 38.2 ± 0.5 | 46.3 ± 0.6 | 2.8 | |
| S14A | * | 8.25 ± 1.02 | 261.9 ± 8.7 | 31.82 ± 4.11 | 40.2 ± 0.3 | 44.1 ± 0.0 | — | ||
| T15A | * | 5.30 ± 0.48 | 504.4 ± 13.0 | 95.25 ± 8.88 | 39.2 ± 0.2 | 42.0 ± 0.0 | — | ||
| S16A | * | 14.01 ± 0.40 | 125.6 ± 0.0 | 8.88 ± 0.22 | 39.4 ± 0.3 | 43.9 ± 0.0 | — | ||
| S16N | — | — | — | — | — | — | — | ||
| S17A | * | 18.45 ± 3.72 | 694.9 ± 62.8 | 37.67 ± 8.23 | 41.7 ± 0.1 | — | — | ||
| 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 | * | — | — | — | — | — | — | ||
| Q19C | * | — | — | — | — | — | — | ||
| ► | Q19M | 0.43 | 0.00 ± 0.03 | 2.9 ± 0.4 | 2,964.50 ± 95,063.40 | 30.0 ± 18.8 | 46.4 ± 1.5 | -18.7 | |
| Q19N | — | — | — | — | — | — | 3.9 | ||
| Q19P | — | — | — | — | — | — | — | ||
| Q19S | * | — | — | — | 36.8 ± 0.2 | 43.1 ± 0.0 | — | ||
| Q19T | 2.19 | 1.74 ± 0.83 | 0.1 ± 0.0 | 0.06 ± 0.03 | — | — | -0.4 | ||
| I20N | 0.31 | 19.41 ± 4.87 | 44.5 ± 4.0 | 2.29 ± 0.61 | — | — | — | ||
| ► | D25Q | 2.73 ± 2.96 | 3.09 ± 0.29 | 292.9 ± 7.2 | 90.27 ± 8.49 | 35.1 ± 287,764,175.7 | — | — | |
| 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 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| D35W | 0.46 | 2.80 ± 0.55 | 13.0 ± 0.6 | 4.66 ± 0.94 | — | — | 14.7 | ||
| F37N | 0.26 | 4.88 ± 0.15 | 8.3 ± 0.1 | 1.70 ± 0.06 | — | — | — | ||
| 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 | — | — | ||
| V52D | 0.20 | 4.76 ± 1.88 | 259.6 ± 16.1 | 54.55 ± 21.78 | — | — | -2.6 | ||
| V52G | * | 8.25 ± 0.54 | 562.9 ± 10.8 | 68.19 ± 4.55 | 39.3 ± 0.1 | — | — | ||
| F57Y | 2.75 | 5.11 ± 0.60 | 37.3 ± 1.1 | 7.29 ± 0.88 | — | 45.2 ± 0.1 | -0.1 | ||
| H58R | 1.22 | 5.64 ± 0.68 | 665.5 ± 21.2 | 118.04 ± 14.68 | — | — | — | ||
| ► | H59K | 1.50 | 2.08 ± 0.82 | 6,326.0 ± 604.1 | 3,046.47 ± 1,233.53 | 41.7 ± 0.5 | — | 1.8 | |
| H59R | 1.35 | 4.83 ± 0.35 | 576.2 ± 10.7 | 119.32 ± 8.87 | — | — | — | ||
| K61S | 0.45 | 4.32 ± 0.84 | 142.2 ± 7.2 | 32.91 ± 6.62 | — | 46.2 ± 0.3 | 0.0 | ||
| 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 | 30.0 ± 96.9 | — | — | ||
| ► | 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 | — | — | — | — | — | — | ||
| D63R | 0.13 | — | — | 0.02 ± 0.00 | — | — | — | ||
| D63S | 0.35 | 0.09 ± 0.16 | 0.1 ± 0.0 | 1.39 ± 2.61 | — | — | — | ||
| K68H | 2.79 | 3.83 ± 0.38 | 397.6 ± 10.1 | 103.74 ± 10.74 | — | 46.8 ± 1.1 | 2.4 | ||
| F72A | * | 5.47 ± 0.28 | 502.2 ± 6.5 | 91.79 ± 4.76 | 38.6 ± 0.4 | — | — | ||
| Y75F | 0.11 | 0.44 ± 0.49 | 1.5 ± 0.3 | 3.51 ± 3.96 | 40.1 ± 1.7 | — | 2.0 | ||
| Y75N | 0.20 | 7.88 ± 0.55 | 410.7 ± 8.0 | 52.13 ± 3.77 | — | — | — | ||
| R76A | — | — | — | — | — | — | — | ||
| R76L | 0.54 | 43.36 ± 1.82 | 7.4 ± 0.1 | 0.17 ± 0.01 | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| R76Q | 0.70 | 6.62 ± 2.33 | 0.8 ± 0.1 | 0.13 ± 0.05 | — | — | -0.1 | ||
| 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 | — | — | |
| ► | A80P | 0.30 | 0.17 ± 0.11 | 0.5 ± 0.1 | 2.94 ± 1.92 | — | 30.6 ± 10.1 | 190.9 | |
| ► | A80S | 1.57 ± 0.69 | 7.92 ± 0.74 | 1,146.2 ± 36.6 | 141.60 ± 12.37 | 40.5 ± 0.5 | 41.1 ± 0.4 | 2.7 | |
| 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 | — | — | ||
| I91H | 3.69 | 11.59 ± 0.81 | 181.2 ± 4.1 | 15.64 ± 1.15 | 37.6 ± 221,464.0 | — | — | ||
| 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 | 0.90 | 8.45 ± 2.50 | 207.6 ± 18.4 | 24.56 ± 7.59 | 36.2 ± 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 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| M114S | 1.18 | 13.79 ± 1.07 | 257.2 ± 6.8 | 18.65 ± 1.53 | — | — | 5.0 | ||
| T116I | 3.42 | 29.39 ± 2.92 | 261.6 ± 10.7 | 8.90 ± 0.95 | — | 42.6 ± 0.5 | -7.3 | ||
| ► | 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 | — | — | ||
| 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 | — | ||
| H119T | 0.84 | 66.09 ± 31.10 | 20.7 ± 5.2 | 0.31 ± 0.17 | — | — | — | ||
| H119V | 0.43 | 44.36 ± 5.88 | 5.2 ± 0.3 | 0.12 ± 0.02 | — | — | — | ||
| W120A | — | — | — | — | — | — | — | ||
| W120D | 0.18 | 12.34 ± 0.44 | 47.0 ± 0.5 | 3.81 ± 0.14 | — | — | — | ||
| 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 | — | — | — | ||
| W120M | 1.46 | 50.86 ± 3.20 | 100.2 ± 3.2 | 1.97 ± 0.14 | — | 43.9 ± 0.1 | 5.2 | ||
| W120N | 0.89 | 0.71 ± 0.29 | 3.6 ± 0.3 | 5.01 ± 2.11 | — | 48.2 ± 0.1 | 4.6 | ||
| W120P | 1.58 | 8.47 ± 0.24 | 1,210.9 ± 9.9 | 143.04 ± 4.25 | — | — | — | ||
| W120Q | 0.63 | — | — | 0.29 ± 0.02 | — | — | — | ||
| W120T | 0.46 | 0.37 ± 0.25 | 0.4 ± 0.0 | 1.01 ± 0.69 | — | 47.0 ± 2.1 | -0.2 | ||
| W120Y | 2.37 | 24.12 ± 3.29 | 1.6 ± 0.1 | 0.07 ± 0.01 | — | 50.2 ± 1.2 | 11.1 | ||
| D121F | — | — | — | — | — | — | — | ||
| Q124I | 1.74 | 3.43 ± 0.22 | 152.5 ± 2.3 | 44.49 ± 2.96 | 40.4 ± 0.7 | — | 5.4 | ||
| 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 | 1.30 | 5.65 ± 0.46 | 181.6 ± 4.0 | 32.13 ± 2.69 | 39.7 ± 0.8 | — | 7.5 | ||
| 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 | — | — | ||
| E129Q | 0.90 | 5.17 ± 0.46 | 348.9 ± 8.3 | 67.49 ± 6.21 | — | 43.4 ± 0.0 | -1.1 | ||
| ► | Q139K | 0.43 ± 0.06 | 6.07 ± 0.79 | 60.1 ± 4.0 | 10.75 ± 3.00 | — | 45.2 ± 0.1 | -2.7 | |
| K142R | 0.54 | 4.57 ± 1.26 | 60.2 ± 4.3 | 13.17 ± 3.74 | — | — | — | ||
| 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 | — | — | ||
| E154A | 0.27 | 30.92 ± 13.46 | 112.2 ± 20.0 | 3.63 ± 1.71 | 30.0 ± 39.3 | — | 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 | — | ||
| 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 | 1.85 ± 0.69 | 6.83 ± 1.28 | 509.8 ± 29.4 | 66.97 ± 13.55 | 38.8 ± 145,420.8 | — | — | |
| 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 | — | — | — | — | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| N163G | 2.38 | 9.68 ± 0.47 | 2.4 ± 0.0 | 0.25 ± 0.01 | — | 41.7 ± 0.0 | 6.0 | ||
| 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 | ||
| 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 | 2.33 | 37.36 ± 2.90 | 487.4 ± 16.6 | 13.05 ± 1.11 | — | — | — | ||
| 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 | |
| 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 | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| S169N | 0.28 | 10.12 ± 5.32 | 1.6 ± 0.2 | 0.16 ± 0.09 | — | — | — | ||
| ► | I170V | 1.22 | 10.02 ± 1.24 | 33,748.0 ± 1,296.6 | 3,368.93 ± 435.92 | 50.0 ± 195,726,000.0 | — | 0.9 | |
| 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 | 2.70 | 1.59 ± 0.12 | 261.4 ± 4.4 | 164.73 ± 12.91 | — | 47.0 ± 0.0 | 1.6 | ||
| L171I | 0.69 | 18.69 ± 3.62 | 427.8 ± 29.3 | 22.89 ± 4.70 | — | — | — | ||
| L171Q | 1.12 | 7.99 ± 0.66 | 77.1 ± 1.8 | 9.66 ± 0.83 | — | — | — | ||
| L171R | * | 3.36 ± 0.23 | 331.2 ± 6.5 | 98.50 ± 6.93 | 38.8 ± 0.3 | 44.6 ± 0.0 | — | ||
| L171Y | 0.91 | 1.10 ± 0.12 | 410.4 ± 9.4 | 373.98 ± 42.07 | 37.2 ± 0.1 | — | — | ||
| 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 | 43.8 ± 4.5 | — | 10.0 | |
| T175K | 2.03 | 6.21 ± 0.54 | 708.5 ± 16.7 | 114.14 ± 10.27 | — | — | — | ||
| T175R | * | 3.59 ± 0.15 | 655.9 ± 6.5 | 182.71 ± 7.79 | 38.1 ± 0.1 | — | — | ||
| E177A | * | 5.98 ± 0.22 | 807.5 ± 8.7 | 135.09 ± 5.20 | 37.5 ± 0.2 | — | — | ||
| 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 | 2.73 | 8.71 ± 0.59 | 504.9 ± 9.9 | 57.94 ± 4.08 | — | — | — | ||
| H178A | * | 7.67 ± 0.73 | 93.1 ± 2.2 | 12.12 ± 1.08 | 39.6 ± 0.4 | — | — | ||
| ► | H178C | 17.97 ± 17.93 | 0.98 ± 0.24 | 36.8 ± 3.8 | 37.27 ± 17.39 | 50.0 ± 975,144,000.0 | — | 3.5 | |
| H178K | 0.67 | 25.15 ± 1.69 | 22.5 ± 0.6 | 0.90 ± 0.06 | — | — | 8.9 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | H178N | 1.16 ± 1.06 | 4.80 ± 0.68 | 340.4 ± 14.2 | 107.79 ± 42.16 | 44.7 ± 4.2 | — | 3.3 | |
| ► | 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 | |
| ► | H182R | 6.90 ± 6.22 | 8.28 ± 0.73 | 604.2 ± 15.8 | 73.34 ± 5.95 | — | — | — | |
| 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 | — | — | — | ||
| 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 | 0.29 | 3.91 ± 0.45 | 130.5 ± 3.8 | 33.40 ± 3.98 | 37.4 ± 0.2 | — | — | ||
| H194Q | 0.16 | 6.08 ± 0.86 | 78.8 ± 3.1 | 12.96 ± 1.90 | 37.2 ± 0.3 | — | — | ||
| ► | H194Y | 0.75 ± 0.01 | 5.62 ± 0.80 | 186.8 ± 6.7 | 33.86 ± 4.11 | 38.9 ± 0.3 | — | — | |
| H206R | 19.65 | 3.12 ± 0.25 | 150.0 ± 3.8 | 48.07 ± 4.02 | — | — | — | ||
| ► | T212E | 1.21 ± 0.07 | 4.93 ± 0.62 | 384.1 ± 13.2 | 77.91 ± 10.34 | — | 46.5 ± 0.2 | -0.2 | |
| T218A | * | 6.51 ± 0.94 | 381.0 ± 15.2 | 58.45 ± 8.66 | 40.1 ± 0.1 | 47.5 ± 0.0 | — | ||
| T218Q | 1.04 | — | — | 0.06 ± 0.00 | — | — | — | ||
| T218S | 1.11 | 14.39 ± 2.25 | 378.3 ± 20.2 | 26.28 ± 4.34 | 41.0 ± 0.2 | — | — | ||
| L219Q | 1.53 | 7.15 ± 0.16 | 1,266.0 ± 7.8 | 177.11 ± 4.03 | — | — | — | ||
| ► | N220A | 0.59 | 10.75 ± 0.65 | 232.1 ± 4.9 | 22.05 ± 1.61 | 38.8 ± 0.2 | 43.6 ± 0.3 | 1.7 | |
| N220C | 1.39 | 1.76 ± 0.46 | 380.3 ± 21.9 | 215.89 ± 57.83 | — | — | — | ||
| N220D | 1.16 | 1.48 ± 0.30 | 100.3 ± 4.5 | 67.69 ± 14.11 | 35.0 ± 0.2 | — | — | ||
| ► | N220F | 2.03 ± 0.74 | 1.42 ± 0.23 | 498.4 ± 11.4 | 551.31 ± 49.53 | 37.9 ± 15,536.4 | — | -2.0 | |
| N220G | * | 33.30 ± 2.36 | 171.0 ± 4.3 | 5.20 ± 0.43 | 41.9 ± 0.2 | 47.4 ± 0.0 | — | ||
| N220H | * | 5.14 ± 0.21 | 101.7 ± 0.0 | 19.70 ± 0.87 | 39.6 ± 0.4 | 46.8 ± 0.0 | — | ||
| N220K | 3.56 | 2.21 ± 0.41 | 351.6 ± 15.0 | 159.19 ± 30.49 | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | N220L | 2.70 ± 1.82 | 13.83 ± 3.62 | 324.7 ± 24.6 | 39.66 ± 10.68 | 36.7 ± 1.2 | — | — | |
| N220Q | 0.29 | 4.46 ± 1.16 | 123.6 ± 9.0 | 27.72 ± 7.52 | 45.7 ± 4.0 | — | 2.6 | ||
| N220R | * | 12.50 ± 1.39 | 13.0 ± 0.0 | 1.08 ± 0.22 | 39.5 ± 0.3 | 46.9 ± 0.0 | — | ||
| N220S | 0.53 | — | — | 6.00 ± 0.19 | 39.7 ± 0.2 | — | — | ||
| ► | N220W | 0.69 ± 0.11 | 0.84 ± 0.22 | 165.3 ± 11.3 | 169.64 ± 46.01 | — | — | — | |
| ► | N220Y | 3.23 | 3.15 ± 0.14 | 481.6 ± 4.2 | 681.98 ± 46.27 | 35.9 ± 17.1 | 49.2 ± 0.0 | — | |
| 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 | 1.03 ± 1.07 | 5.10 ± 1.29 | 427.9 ± 40.6 | 78.89 ± 19.79 | 42.8 ± 2.7 | 41.6 ± 0.3 | -2.5 | |
| P231K | 2.98 | 4.57 ± 0.42 | 349.8 ± 8.5 | 76.53 ± 7.33 | — | 44.8 ± 0.2 | 0.2 | ||
| A236D | — | — | — | — | — | — | 43.0 | ||
| A236E | — | — | — | — | — | — | — | ||
| ► | A236V | 1.03 ± 0.52 | 0.45 ± 0.31 | 1.2 ± 0.2 | 2.51 ± 1.90 | 36.2 ± 1.3 | — | — | |
| I238K | 1.76 | 9.56 ± 1.74 | 2,004.1 ± 112.1 | 209.71 ± 39.95 | 39.4 ± 0.2 | — | — | ||
| ► | R240A | 2.57 ± 1.04 | 11.85 ± 2.51 | 419.3 ± 20.1 | 35.94 ± 5.10 | 38.5 ± 0.2 | — | 2.3 | |
| R240D | * | 10.82 ± 0.47 | 231.6 ± 4.3 | 21.43 ± 1.08 | 37.2 ± 0.0 | 45.7 ± 0.0 | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| R240E | — | — | — | — | — | — | — | ||
| R240G | 0.30 | 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 | — | — | — | ||
| ► | F243H | 1.15 ± 0.43 | 21.06 ± 6.11 | 552.5 ± 64.9 | 36.16 ± 5.42 | 39.4 ± 0.3 | — | 1.4 | |
| 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 | — | — | — | ||
| 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 | — | — | — | — | — | — | — | ||
| F253H | 0.28 | 64.81 ± 42.05 | 297.9 ± 110.9 | 4.60 ± 3.44 | — | — | 3.6 | ||
| ► | F253I | 2.66 ± 1.40 | 5.68 ± 0.58 | 442.8 ± 12.3 | 78.01 ± 8.21 | — | 44.8 ± 0.1 | 5.9 | |
| ► | F253M | 2.17 ± 1.17 | 5.99 ± 0.39 | 448.5 ± 8.2 | 37.49 ± 3.61 | — | 44.7 ± 0.1 | 6.8 | |
| F253S | 0.82 | 1.65 ± 0.23 | 11,626.5 ± 341.4 | 7,052.98 ± 1,010.15 | 39.5 ± 0.4 | — | — | ||
| F253Y | 0.23 | 6.14 ± 0.61 | 271.2 ± 7.5 | 44.21 ± 4.56 | — | — | 1.4 | ||
| Y257F | 1.59 | 1.86 ± 0.67 | 45.4 ± 3.5 | 24.42 ± 9.03 | 38.3 ± 0.9 | 58.2 ± 30.6 | 1.1 | ||
| 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 | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| 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 | ||
| L269I | 0.21 | 3.77 ± 0.53 | 176.9 ± 6.2 | 46.90 ± 6.80 | 40.4 ± 0.6 | — | 0.4 | ||
| 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 | 2.29 | 5.38 ± 0.48 | 772.8 ± 18.0 | 143.61 ± 13.13 | — | — | — | ||
| ► | 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 | |
| N293C | * | 19.80 ± 2.08 | 8.7 ± 0.0 | 0.43 ± 0.00 | 38.9 ± 0.4 | 47.0 ± 0.0 | — | ||
| N293D | — | — | — | — | — | — | — | ||
| ► | N293G | 3.88 ± 2.24 | 24.01 ± 2.80 | 1.1 ± 0.0 | 0.04 ± 0.01 | — | — | — | |
| N293K | — | — | — | — | — | — | — | ||
| N293M | * | — | — | — | — | — | 41.9 | ||
| N293Q | * | — | — | — | — | — | — | ||
| N293R | 0.37 | 0.70 ± 0.15 | 7.9 ± 0.3 | 11.25 ± 2.44 | — | — | — | ||
| N293S | 2.91 | 0.45 ± 0.18 | 2.2 ± 0.2 | 4.91 ± 1.96 | — | 47.8 ± 0.5 | 0.5 | ||
| ► | N293V | 1.64 ± 0.40 | 6.80 ± 0.91 | 0.1 ± 0.0 | 0.01 ± 0.00 | 37.9 ± 67,349.9 | — | — | |
| N293W | — | — | — | — | — | — | 132.7 | ||
| N293Y | 0.61 | 48.85 ± 36.83 | 0.6 ± 0.2 | 0.01 ± 0.01 | — | 49.0 ± 0.8 | — | ||
| 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 | — | ||
| Y294L | — | — | — | — | — | — | — | ||
| Y295A | * | — | — | — | — | — | — | ||
| Y295G | * | — | — | — | — | — | — | ||
| Y295N | 1.59 | 18.93 ± 3.85 | 140.3 ± 10.5 | 7.41 ± 1.61 | — | — | — | ||
| ► | T296A | 2.99 | 47.09 ± 4.63 | 0.6 ± 0.0 | 0.01 ± 0.00 | — | — | — | |
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| ► | T296C | 0.60 ± 0.43 | 10.65 ± 1.48 | 248.3 ± 15.2 | 21.43 ± 4.20 | — | — | — | |
| T296E | — | — | — | — | — | — | — | ||
| T296H | 0.46 | 8.23 ± 1.10 | 101.5 ± 3.9 | 12.33 ± 1.72 | — | — | — | ||
| T296Y | 4.35 | 5.95 ± 0.52 | 497.2 ± 12.0 | 83.60 ± 7.60 | — | 45.4 ± 0.0 | -1.1 | ||
| 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 | 7.15 ± 5.51 | 5.53 ± 0.32 | 2,213.5 ± 43.4 | 400.47 ± 30.68 | — | — | — | |
| ► | S302H | 3.21 ± 2.02 | 4.81 ± 0.37 | 568.3 ± 14.0 | 133.50 ± 16.70 | — | — | — | |
| S302N | 3.27 | 5.43 ± 0.16 | 718.7 ± 5.4 | 132.24 ± 3.92 | — | — | — | ||
| A306K | 1.55 | 4.71 ± 0.54 | 698.6 ± 21.3 | 148.21 ± 17.68 | 40.7 ± 0.2 | — | -1.5 | ||
| ► | A306Q | 2.08 | 3.75 ± 0.63 | 587.8 ± 25.0 | 156.87 ± 27.23 | 38.3 ± 0.8 | — | — | |
| ► | A306S | 1.26 ± 0.63 | 9.48 ± 1.00 | 460.1 ± 13.8 | 51.42 ± 5.25 | 40.4 ± 0.1 | 43.7 ± 0.1 | -1.5 | |
| V311D | 3.05 | 6.84 ± 0.65 | 295.0 ± 8.0 | 43.14 ± 4.25 | — | 41.8 ± 0.1 | 2.6 | ||
| 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 | — | — | — | ||
| H315D | 0.32 | 0.68 ± 0.24 | 897.0 ± 54.3 | 1,312.84 ± 460.56 | 40.1 ± 0.1 | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| H315E | * | — | — | — | — | — | — | ||
| 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 | 0.79 | 4.27 ± 2.31 | 63.5 ± 9.4 | 14.89 ± 8.35 | 50.0 ± 0.0 | — | -1.5 | |
| H315P | 0.84 | 5.39 ± 0.27 | 647.6 ± 9.8 | 120.14 ± 6.24 | 38.8 ± 0.2 | — | -1.8 | ||
| H315T | 4.76 | 4.67 ± 0.11 | 658.5 ± 4.0 | 141.04 ± 3.48 | — | — | — | ||
| 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 | — | — | — | ||
| 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 | 5.27 | 8.15 ± 0.35 | 344.0 ± 4.4 | 42.22 ± 1.90 | — | 44.9 ± 0.1 | 6.4 | ||
| M323K | — | — | — | — | — | — | — | ||
| W325A | * | 1.61 ± 0.23 | 23.8 ± 0.0 | 14.72 ± 2.16 | 40.1 ± 0.4 | 41.8 ± 0.0 | — | ||
| 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 | — | — | — | ||
| W325F | 1.87 | 12.63 ± 3.27 | 116.0 ± 9.7 | 9.19 ± 2.50 | — | — | — | ||
| W325G | — | — | — | — | — | — | — | ||
| W325H | * | 3.08 ± 0.43 | 28.1 ± 0.0 | 9.09 ± 1.30 | 37.7 ± 0.3 | 40.7 ± 0.0 | — | ||
| ► | W325K | 0.54 ± 0.22 | 3.58 ± 0.97 | 4.5 ± 0.4 | 0.92 ± 0.37 | 50.0 ± 11.9 | — | 3.6 | |
| W325L | * | 5.74 ± 0.35 | 88.8 ± 2.2 | 15.37 ± 1.08 | 40.4 ± 0.5 | — | — | ||
| ► | W325M | 3.03 ± 1.36 | 5.83 ± 0.56 | 52.0 ± 1.3 | 8.91 ± 1.03 | — | 42.8 ± 0.4 | 3.5 | |
| ► | W325R | 2.05 | 14.18 ± 3.12 | 213.4 ± 15.7 | 15.05 ± 3.49 | 40.0 ± 0.2 | 42.7 ± 0.0 | — | |
| ► | H328K | 1.65 ± 0.05 | 7.30 ± 2.91 | 338.8 ± 42.9 | 91.26 ± 33.42 | 38.8 ± 0.3 | — | 1.1 | |
| H328N | 0.64 | 0.79 ± 0.36 | 3,454.4 ± 308.9 | 4,359.60 ± 2,002.14 | 37.2 ± 0.8 | — | 0.6 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| 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 | — | — | — | ||
| ► | Y333F | 2.28 ± 0.52 | 1.93 ± 0.20 | 352.2 ± 11.0 | 192.18 ± 28.53 | 37.8 ± 59,885.7 | — | — | |
| 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 | 1.85 ± 0.85 | 1.59 ± 0.22 | 408.6 ± 14.9 | 311.92 ± 110.17 | 38.3 ± 0.2 | — | -6.8 | |
| 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 | |
| F343S | — | — | — | — | — | — | — | ||
| L350I | 1.67 | 5.71 ± 0.48 | 535.9 ± 11.9 | 93.83 ± 8.09 | — | — | — | ||
| I351V | 3.95 | 4.78 ± 0.39 | 436.1 ± 9.2 | 91.20 ± 7.74 | — | — | — | ||
| ► | T352A | 0.03 | 14.26 ± 1.76 | 49.8 ± 2.2 | 3.46 ± 0.43 | 38.1 ± 0.3 | 39.3 ± 0.0 | 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 | 50.0 ± 12.1 | — | — | |
| T352W | — | — | — | — | — | — | 71.5 | ||
| E353A | * | — | — | — | — | — | — | ||
| E353D | 0.45 | 2.35 ± 0.99 | 1.6 ± 0.2 | 0.68 ± 0.29 | 50.0 ± 10.9 | — | — | ||
| N354A | * | 5.38 ± 0.67 | 2.2 ± 0.0 | 0.43 ± 0.00 | 40.8 ± 0.4 | 46.7 ± 0.0 | — | ||
| G355A | — | — | — | — | — | — | — | ||
| G355N | 0.31 | 8.65 ± 2.96 | 0.4 ± 0.0 | 0.04 ± 0.01 | — | — | -3.6 | ||
| 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 | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| 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 | — | — | — | ||
| 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 | — | — | — | ||
| 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 | 1.05 | 5.02 ± 0.49 | 35.3 ± 0.9 | 7.03 ± 0.70 | — | — | -1.1 | ||
| H384A | 1.09 | 6.22 ± 0.42 | 644.9 ± 11.9 | 103.69 ± 7.30 | — | — | — | ||
| 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 | ||
| ► | E388L | 3.46 ± 0.33 | 5.29 ± 0.90 | 136.5 ± 3.8 | 30.90 ± 3.17 | 40.2 ± 0.2 | — | — | |
| E388R | 4.14 | 3.67 ± 0.80 | 304.6 ± 16.4 | 82.96 ± 18.62 | — | — | — | ||
| 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 | ||
| ► | Y396P | — | — | — | — | — | — | 283.8 | |
| Y396S | — | — | — | — | — | — | 8.2 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| Y396V | — | — | — | — | — | — | 21.5 | ||
| Y396W | 1.28 | 1.96 ± 0.54 | 14.1 ± 1.1 | 7.20 ± 2.05 | 38.4 ± 0.5 | — | 3.9 | ||
| 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 | * | — | — | — | — | — | — | ||
| W399R | * | 12.75 ± 0.87 | — | — | 37.9 ± 0.8 | 49.1 ± 0.0 | — | ||
| 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 | ||
| 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 | 1.19 | 49.80 ± 3.48 | 18.7 ± 0.7 | 0.38 ± 0.03 | — | — | — | ||
| ► | N404E | 1.69 ± 1.63 | 5.66 ± 0.89 | 66.8 ± 3.5 | 18.87 ± 4.15 | 40.0 ± 0.4 | 45.3 ± 0.6 | 3.9 | |
| ► | N404L | 1.90 ± 0.75 | 30.74 ± 2.08 | 22.8 ± 0.6 | 0.74 ± 0.05 | — | 44.6 ± 0.1 | -1.6 | |
| ► | N404M | 2.17 ± 1.10 | 21.49 ± 2.36 | 18.1 ± 1.7 | 1.54 ± 0.54 | 40.6 ± 0.3 | 48.4 ± 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 | 4.96 | 23.55 ± 5.86 | 23.4 ± 2.2 | 0.99 ± 0.26 | — | — | — | ||
| 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 | — | — | — | — | — | — | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| E406H | 2.36 | 0.02 ± 0.04 | 0.0 ± 0.0 | 1.80 ± 2.83 | — | — | 4.5 | ||
| ► | E406Q | 2.98 ± 1.48 | 53.64 ± 4.27 | 7.3 ± 0.3 | 0.14 ± 0.01 | — | 44.3 ± 0.5 | 4.3 | |
| E406T | 3.40 | 2.57 ± 1.38 | 15.6 ± 1.9 | 6.05 ± 3.33 | — | — | — | ||
| W407A | * | — | — | — | — | — | — | ||
| W407D | 0.14 | 0.37 ± 0.90 | 3.0 ± 1.4 | 8.05 ± 20.19 | — | — | 2.5 | ||
| W407F | 0.98 | 38.81 ± 4.02 | 6.1 ± 0.3 | 0.16 ± 0.02 | — | 44.8 ± 0.0 | 6.1 | ||
| 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 | — | — | — | |
| W407Q | — | — | — | — | — | — | — | ||
| ► | W407R | — | — | — | — | — | — | 8.8 | |
| W407Y | — | — | — | — | — | — | — | ||
| A408F | 0.31 | 37.97 ± 8.36 | 341.9 ± 33.2 | 9.00 ± 2.17 | — | — | — | ||
| A408K | 2.59 | 6.32 ± 0.92 | 319.8 ± 12.7 | 50.56 ± 7.62 | — | — | — | ||
| 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 | ||
| W409D | 3.60 | 2.29 ± 0.14 | 532.9 ± 7.4 | 233.16 ± 14.49 | — | — | — | ||
| ► | W409E | 1.39 ± 0.85 | 2.11 ± 0.16 | 636.6 ± 9.5 | 297.60 ± 17.79 | — | 46.8 ± 0.1 | 4.9 | |
| W409F | 0.78 | 3.65 ± 0.29 | 615.2 ± 12.3 | 168.53 ± 13.79 | 39.9 ± 0.1 | — | — | ||
| W409G | 0.44 | 3.85 ± 0.82 | 14.1 ± 0.7 | 3.66 ± 0.81 | — | — | — | ||
| W409N | 4.93 | 8.43 ± 1.13 | 298.7 ± 12.0 | 35.42 ± 4.97 | — | 38.9 ± 0.1 | 1.9 | ||
| ► | W409R | 2.67 ± 0.72 | 2.73 ± 0.32 | 142.5 ± 4.1 | 52.07 ± 6.36 | — | 44.5 ± 0.3 | 3.7 | |
| ► | W409Y | 0.26 | 2.79 ± 0.29 | 693.8 ± 20.3 | 248.84 ± 25.30 | 36.8 ± 0.1 | 43.5 ± 0.0 | 0.4 | |
| ► | Y411R | 0.28 ± 0.13 | 1.99 ± 0.63 | 50.3 ± 3.3 | 32.27 ± 11.15 | — | — | — | |
| K413Q | 3.02 | 5.41 ± 0.62 | 386.6 ± 12.1 | 71.53 ± 8.55 | — | 45.8 ± 1.7 | 5.0 | ||
| Variant | Yield, cE (mg/mL) |
KM (mm) | kcat (min−1) |
kcat/KM (mm−1min−1) |
T50 (°C) | Tm (°C) | Rosetta score change |
||
| K413R | 0.41 | 0.85 ± 0.29 | 470.8 ± 32.0 | 555.04 ± 196.18 | 40.1 ± 0.1 | — | — | ||
| F415A | * | 16.63 ± 4.00 | — | — | 40.5 ± 0.3 | 44.7 ± 0.0 | — | ||
| F415N | * | — | — | — | — | — | — | ||
| F415Y | 2.24 | 10.01 ± 1.13 | 288.3 ± 9.8 | 28.82 ± 3.40 | — | — | — | ||
| ► | H419Y | 3.27 ± 0.04 | 4.88 ± 0.44 | 742.6 ± 14.1 | 143.16 ± 11.99 | — | 49.1 ± 1.1 | -2.1 | |
| 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 | — | ||
| R427H | 0.46 | 1.28 ± 0.41 | 1.0 ± 0.1 | 0.80 ± 0.26 | — | — | 11.6 | ||
| ► | R427L | 0.41 | 4.95 ± 0.40 | 380.1 ± 8.3 | 76.73 ± 6.49 | — | 45.4 ± 0.3 | 4.9 | |
| R427W | 0.31 | 0.98 ± 0.27 | 35.6 ± 2.0 | 36.11 ± 10.00 | 40.2 ± 0.1 | — | — | ||
| S432A | 1.12 | 2.23 ± 0.34 | 69.0 ± 2.3 | 30.97 ± 4.86 | — | 79.3 ± 0.0 | 0.5 | ||
| L434S | 2.49 | 0.57 ± 0.16 | 239.0 ± 13.1 | 416.14 ± 121.04 | — | 43.2 ± 0.0 | 1.2 | ||
| W435Y | 0.48 | 7.41 ± 0.72 | 823.9 ± 23.2 | 111.17 ± 11.22 | — | — | — | ||
| A441V | 2.07 | 4.65 ± 0.10 | 859.5 ± 4.6 | 184.98 ± 4.01 | — | — | — |