Table 2.
HsKYNase variants critical in the evolution of HsKYNase_93D9 and their respective amino acid substitutions
| Enzyme species | Type of mutagenesis | # of mutations | Amino acid substitutions | Fold relative to HsKYNase |
|---|---|---|---|---|
| HsKYNase_1 | NNS combinatorial mutagenesis | 4 | aF71M-L72N-K81R-M82N | 17.2 |
| HsKYNase_8 | NNS combinatorial mutagenesis | 3 | E103F-V104H-G105T | 17 |
| Combo variant_1 | Rational combination of beneficial mutations | 6 | F71M-L72N-K81R-M82N-I331N-F306Y | 16 |
| Rationally-designed variant_12 | Rational combination of beneficial mutations | 6 | E259P-W272G-S274G-A280A-A282P-G287F | 3 |
| B-factor_10 | NNS & rational combination of beneficial mutations | 7 | I183P-E184A-M187L-M189I-K191N-E197T-I201T | 6 |
| HsKYNase_93 | DNA shuffling | 15 | F71M-L72N-E103F-V104H-G105T-I183P-E184A-M187L-M189I-K191N-E197T-I201T-A280S-A282P-F306Y | 131 |
| HsKYNase_93D9 | NNS combinatorial mutagenesis & DNA shuffling | 17 |  |
170 |
aMutations that are incorporated in HsKYNase_93 through DNA shuffling are shown in underlined and bold font throughout the Table.
bThe two additional amino acid substitutions of HsKYNase_93D9 relative to HsKYNase_93 are shown in blue. All mutations in bold font are present in both HsKYNase_93 and HsKYNase_93D9.