9H87
Crystal structure of LmrR variant V15aY with Val15 replaced by 3-aminotyrosine
Summary for 9H87
| Entry DOI | 10.2210/pdb9h87/pdb |
| Descriptor | Transcriptional regulator, PadR-like family (2 entities in total) |
| Functional Keywords | artificial enzyme, friedel-crafts alkylase, unnatural amino acid, 3-aminotyrosine, dna binding protein |
| Biological source | Lactococcus cremoris subsp. cremoris MG1363 |
| Total number of polymer chains | 1 |
| Total formula weight | 15069.85 |
| Authors | Thunnissen, A.M.W.H.,Brouwer, B.,Roelfes, G. (deposition date: 2024-10-28, release date: 2025-04-30, Last modification date: 2025-05-07) |
| Primary citation | Brouwer, B.,Della-Felice, F.,Thunnissen, A.W.H.,Roelfes, G. Genetically encoded 3-aminotyrosine as catalytic residue in a designer Friedel-Crafts alkylase. Chem Sci, 2025 Cited by PubMed Abstract: Genetic incorporation of noncanonical amino acids (ncAAs) harbouring catalytic side chains into proteins allows the creation of enzymes able to catalyse reactions that have no equivalent in nature. Here, we present for the first time the use of the ncAA 3-aminotyrosine (aY) as catalytic residue in a designer enzyme for iminium activation catalysis. Incorporation of aY into protein scaffold LmrR gave rise to an artificial Friedel-Crafts (FC) alkylase exhibiting complementary enantioselectivity to a previous FC-alkylase design using -aminophenylalanine as catalytic residue in the same protein. The new FC-alkylase was optimized by directed evolution to afford a quadruple mutant that showed increased activity and excellent enantioselectivity (up to 95% ee). X-ray crystal structures of the parent and evolved designer enzymes suggest that the introduced mutations cause a narrowing of the active site and a reorientation of the catalytic -NH group. Furthermore, the evolved FC-alkylase was applied in whole-cell catalysis, facilitated by the straightforward incorporation of aY. Our work demonstrates that aY is a valuable addition to the biochemists toolbox for creating artificial enzymes. PubMed: 40276638DOI: 10.1039/d5sc01055a PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.15 Å) |
Structure validation
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