Table 2 Experimental characterization results of the afCR multi-site mutants
Enzyme | Mutation | 57 °C t1/2 (min) | Fold improvement | Relative activity (%) |
|---|---|---|---|---|
M0 | I304L/F395V | 2 | 1 | 100 |
M1 | M0 + D17V | 40 | 20 | 105 ± 7.71 |
M2-1 | M1 + L6P | 142 | 71 | 104.76 ± 18.10 |
M2-2 | M1 + T251C | 71 | 35.5 | 118.09 ± 3.25 |
M2-3 | M1 + K351E | 101 | 50.5 | 161.90 ± 1.90 |
M2-4 | M1 + T199S | 210 | 105 | 111.43 ± 2.31 |
M3-1 | M2-4+ T251C | 599 | 299.5 | 112.38 ± 2.86 |
M3-2 | M2-4+ F108Y | 482 | 241 | 106.67 ± 4.76 |
M3-3 | M2-4+ K351E | 859 | 429.5 | 157.61 ± 5.30 |
M3-4 | M2-4+ L6P | 1258 | 629 | 123.81 ± 1.90 |
M4-1 | M3-4+ F108Y | 2498 | 1249 | 134.28 ± 2.86 |
M4-2 | M3-4+ T251C | 3371 | 1685.5 | 110.47 ± 1.90 |