| Wild Type | A 4.1 | A 4.3 | A 4.5 | A 5.1 | A 5.2 | B 4.1 | C 4.1 | C 4.2 | Neutral 1 | Neutral 2 | Shuffled-1 | Shuffled-2 |
---|
AA substitutions | | 2 | 3 | 4 | 4 | 3 | 2 | 3 | 3 | 2 | 5 | | |
Silent Mutations | | 2 | 1 | 1 | 3 | 0 | 1 | | | | | | |
Position | | | | | | | | | | | | | |
36 | E | | | | | | | | | | K | | |
42 | S | | | | | | | | | | | N | |
57 | D | | | | N | | | | | | | | |
67 | V | | | | | | | | D | | | | |
81 | G | | | | | V | | | | | | | V |
185 | D | | | | | | | | | | G | | |
168 | A | | | | | S | | | | | | | |
217 | R | | | C | | | | | | | | | |
245 | G | | | | | | | A | | | | | |
266 | P | | | | | | | | | | S | | |
278 | G | | A | | | | | | | | | | |
308 | N | | D | | | | | | | | | | |
313 | H | | | | | | | | | | Y | | |
367 | A | | | P | | | | | | | | | |
370 | K | | | | | | | R | R | | | | |
378 | E | | | | | | | | | | | R | |
480 | T | | | | | | A | | | | | | |
484 | V | | | | | | | | | I | | | |
498 | Q | | | | | | | | | | | | R |
508 | D | | | | G | | | | | | | | |
509 | T | | A | A | | | | | | | | A | A |
510 | L | F | | | | | | | | | | | |
529 | W | | | | | L | | L | L | | | | |
533 | Y | | | | C | | | | | | | | |
536 | V | | | | | | | | | I | | | |
540 | V | | | D | | | | | | | | | |
557 | S | | | | A | | | | | | | P | P |
566 | N | S | | | | | S | | | | | S | S |
568 | K | | | | | | | | | | | Q | Q |
594 | G | | | | | | | | | | D | | |
- Variants A4.1 - 4.5 were obtained from experiment A after four rounds of neutral screening. Variants A5.1 and A5.2 were obtained from the fifth round of neutral screening. Variant B4.1 was obtained from experiment B using a high mutagenesis rate, after four rounds of neutral screening. Variants C4.1 and C4.2 were obtained from experiment C after four rounds of neutral screening. Shuffled-1 and Shuffled-2 are the variants obtained by DNA shuffling of the variants with galactosidase activity. The positions where mutations were previously identified by Matsumura and Ellington [13] are highlighted.