Table II.
Summary of enzymes used in the development of date-driven enzyme engineering models for enhanced activity or specificitya
| Enzyme | Substrate | Mutation | Performance | Paper |
|---|---|---|---|---|
| 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylic acid synthase | aldehyde | NA | new substrate specificity | 2017-Pertusi (Pertusi et al., 2017) |
| carboxylic acid reductase | carboxylic acid | NA | new substrate specificity | 2017-Pertusi (Pertusi et al., 2017) |
| amino acid ester hydrolase | amide, ester | NA | new substrate specificity | 2017-Pertusi (Pertusi et al., 2017) |
| 4-hydroxyacetophenone monooxygenase | keton | NA | new substrate specificity | 2017-Pertusi (Pertusi et al., 2017) |
| phosphotriesterase | phosphotriester, ester, lactone | I106L/F132L/H254R/H257W/L303T | 741664-fold specificity enhancement (paraoxon) | 2018- Khersonsky (Khersonsky et al., 2018) |
| acetyl-CoA synthetase | CoA, aliphatic acid | V310I/T311V/S314T/Y355F/V386L/F421A | 7-fold activity enhancement (butyrate) | 2018- Khersonsky (Khersonsky et al., 2018) |
| glycosyltransferase | sugar donor, glycosyl acceptor | NA | new sequence for new substrate | 2018-Yang (Yang et al., 2018) |
| phosphomannose isomerase | phosphomannose | D229W/N272K/L335A/N388S/S425T | 5-fold activity enhancement | 2020-Shroff (Shroff et al., 2020) |
| TEM-1 β-lactamase | carbenicillin | N52, F60, Q88, Q99, T114, M182, E197 | gain of new function | 2020-Shroff (Shroff et al., 2020) |
| 2-deoxy-D-ribose 5-phosphate aldolase | acetaldehyde | C47V/G204A/S239D | ~3-fold activity enhancement (acetaldehyde), abolishment of natural activity (deoxyribose-5-phosphate, deoxyribose) | 2020-Voutilainen (Voutilainen et al., 2020) |
| thiolase | p-nitrophenyl ester | NA | Specificity-determining residue, structural/chemical feature influencing activity | 2020-Robinson (Robinson et al., 2020) |
| nitrilase | nitriles | NA | substrate scope expansion | 2021-Mou (Mou et al., 2021) |
aIn the column of Mutation and Performance, only the best-performing enzyme mutants are shown. Mutations separated by slash ‘/’ indicate multiple mutations in one variant. Mutations separated by comma ‘,’ indicate different variants with single amino acid substitution.