Table 5.
Catalytic performance of RhlAPa variants explored in this study.
| Mutation | Relative activity* | Mutation | Relative activity* | Mutation | Relative activity* |
|---|---|---|---|---|---|
| A36L | 6 | D137A | 32 (± 3) | R169K | 126 (± 10) |
| A36V | 13 (± 2) | D137E | 51 (± 8) | E185A | 12 (± 1) |
| A36M | 43 (± 7) | Y138A | 12 (± 1) | Q188N | 29 (± 3) |
| A36I | 7 | Y138F | 14 (± 1) | A189P | 1 |
| A36G | 2 | R141A | 63 (± 5) | H192R | 1 |
| M37A | 0 | R141A-K164R | 97 (± 5) | H192A | 9 (± 2) |
| M37V | 43 (± 4) | R141K | 121 (± 11) | H192K | 5 (± 1) |
| M37I | 9 (± 1) | R141A-T161A-K164R | 0 | A198P | 10 (± 1) |
| M37L | 123 (± 6) | A142P | 2 | L199A | 0 |
| M37F | 12 (± 1) | A142P-A144P | 2 | L199M | 4 (± 2) |
| Q66R | 68 (± 7) | L145H | 60 (± 4) | L199V | 29 (± 9) |
| Q69K | 8 (± 1) | L145A | 27 (± 4) | L199I | 54 (± 13) |
| Q69N | 138 (± 10) | L145I | 23 (± 2) | R202K | 201 (± 4) |
| Q69R | 0 | L146I | 56 (± 9) | R202E | 0 |
| K79Q | 6 | L148I | 86 (± 13) | R202D | 0 |
| K79R | 9 (± 1) | L157A | 5 | Y204E | 93 (± 7) |
| S102A | 0 | L157I | 18 (± 4) | Y204D | 88 (± 7) |
| W103A | 11 (± 1) | L157P | 2 | D223A | 0 |
| W103Y | 9 (± 1) | L158A | 3 | E224A | 99 (± 18) |
| A128G | 63 (± 14) | L158P | 2 | E224D | 134 (± 2) |
| A128V | 55 (± 8) | L158I | 101 (± 5) | Y225A | 0 |
| A128L | 7 (± 1) | E160A | 2 | Y225F | 14 (± 1) |
| A128I | 5 (± 2) | T161A | 57 (± 3) | Y225S | 6 |
| A128W | 0 | T161S | 23 (± 5) | T227E | 31 (± 2) |
| A128S | 57 (± 7) | K164R-T227S | 60 (± 8) | T227D | 38 (± 2) |
| L131A | 21 (± 2) | K164R-Y165F | 59 (± 6) | T227S | 145 (± 15) |
| L131V | 0 | K164A | 55 (± 10) | T227D | 38 (± 6) |
| L131I | 86 (± 24) | Y165A | 4 | D230E | 148 (± 24) |
| N132K | 3 | Y165F | 187 (± 16) | H251A | 0 |
| N132R | 15 (± 1) | Y165F-T227S | 13 | E256D | 0 |
| N132Q | 11 (± 1) |
*
Catalytic activity is relative to the WT RhlAPa performance with standard deviation (SD) obtained from triplicate experiments.