EC Number |
Protein Variants |
Reference |
---|
2.6.1.B21 | H106Y/Y108R |
site-directed mutagenesis |
-, 758848 |
2.6.1.B21 | H86F |
site-directed mutagenesis, the mutant shows no activity with (R)-PEA and no wild-type DATA activity |
758564 |
2.6.1.B21 | more |
creation of an (R)-amine transaminase activity within an alpha-amino acid transaminase scaffold via one to six amino acid substitutions in the enzyme's active site. The final sextuple variant M2-6 shows almost complete depletion of the native DATA activity but high activity toward (R)-phenylethylamine ((R)-PEA) and has a high specific activity in the conversion of (R)-PEA and pyruvate to acetophenone and D-alanine. The crystal structure of the D-amino acid aminotransferase (DATA, UniProt ID P19938, EC 2.6.1.21) from Bacillus sp. strain YM-1 (PDB entry 3DAA) is used as a scaffold for the computational enzyme redesign. Molecular dynamics simulations and molecular mechanics Poisson-Boltzmann surface area (MM/PBSA) calculations |
758564 |
2.6.1.B21 | more |
mutational analysis of enzyme-substrate interactions and substrate specificity, overview. The evolved ATAs perform asymmetric synthesis of the respective R-amine with high conversions by using either alanine or isopropylamine |
759050 |
2.6.1.B21 | P281S |
site-directed mutagenesis, the mutation seems to have a negative effect on specific activity |
759050 |
2.6.1.B21 | S180A |
site-directed mutagenesis, the mutant shows no activity with (R)-PEA and no wild-type DATA activity |
758564 |
2.6.1.B21 | Y152F |
site-directed mutagenesis, the mutation stabilizes the enzyme, the activity is slightly reduced compared to wild-type |
759050 |
2.6.1.B21 | Y31F |
site-directed mutagenesis, the mutant shows low activity with (R)-PEA and good wild-type DATA activity |
758564 |
2.6.1.B21 | Y31F/H86F |
site-directed mutagenesis, variant M2 exhibited some (R)-PEA acceptance when PMP formation after incubating the purified variant with (R)-PEA is assayed, which indicates a completed first half-reaction. Lack of activity in the acetophenone assay |
758564 |
2.6.1.B21 | Y31F/H86F/S180A/T242I |
site-directed mutagenesis, variant M2-4 shows significant activity toward (R)-PEA, and retains no native DATA activity |
758564 |