9EQ5
CryoEM Structure of Phenylalanine Ammonia Lyase from Planctomyces brasiliencis
Summary for 9EQ5
| Entry DOI | 10.2210/pdb9eq5/pdb |
| EMDB information | 19897 |
| Descriptor | Histidine ammonia-lyase, [(1R)-1-amino-2-phenylethyl]phosphonic acid (2 entities in total) |
| Functional Keywords | phenylalanine catabolism lyase, lyase |
| Biological source | Rubinisphaera brasiliensis |
| Total number of polymer chains | 4 |
| Total formula weight | 251645.97 |
| Authors | Duhoo, Y.,Buslov, I.,Desmons, S. (deposition date: 2024-03-20, release date: 2024-08-07, Last modification date: 2024-10-09) |
| Primary citation | Buslov, I.,Desmons, S.,Duhoo, Y.,Hu, X. Engineered Phenylalanine Ammonia-Lyases for the Enantioselective Synthesis of Aspartic Acid Derivatives. Angew.Chem.Int.Ed.Engl., 63:e202406008-e202406008, 2024 Cited by PubMed Abstract: Biocatalytic hydroamination of alkenes is an efficient and selective method to synthesize natural and unnatural amino acids. Phenylalanine ammonia-lyases (PALs) have been previously engineered to access a range of substituted phenylalanines and heteroarylalanines, but their substrate scope remains limited, typically including only arylacrylic acids. Moreover, the enantioselectivity in the hydroamination of electron-deficient substrates is often poor. Here, we report the structure-based engineering of PAL from Planctomyces brasiliensis (PbPAL), enabling preparative-scale enantioselective hydroaminations of previously inaccessible yet synthetically useful substrates, such as amide- and ester-containing fumaric acid derivatives. Through the elucidation of cryo-electron microscopy (cryo-EM) PbPAL structure and screening of the structure-based mutagenesis library, we identified the key active site residue L205 as pivotal for dramatically enhancing the enantioselectivity of hydroamination reactions involving electron-deficient substrates. Our engineered PALs demonstrated exclusive α-regioselectivity, high enantioselectivity, and broad substrate scope. The potential utility of the developed biocatalysts was further demonstrated by a preparative-scale hydroamination yielding tert-butyl protected l-aspartic acid, widely used as intermediate in peptide solid-phase synthesis. PubMed: 38713131DOI: 10.1002/anie.202406008 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (2.17 Å) |
Structure validation
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