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Title | Engineered Phenylalanine Ammonia-Lyases for the Enantioselective Synthesis of Aspartic Acid Derivatives. |
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Journal, issue, pages | Angew Chem Int Ed Engl, Vol. 63, Issue 31, Page e202406008, Year 2024 |
Publish date | Jul 29, 2024 |
Authors | Ivan Buslov / Sarah Desmons / Yoan Duhoo / Xile Hu / |
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 ...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. |
External links | Angew Chem Int Ed Engl / PubMed:38713131 |
Methods | EM (single particle) |
Resolution | 2.17 Å |
Structure data | EMDB-19897, PDB-9eq5: |
Chemicals | ChemComp-PPH: |
Source |
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Keywords | LYASE / Phenylalanine catabolism Lyase |