9EQ5

CryoEM Structure of Phenylalanine Ammonia Lyase from Planctomyces brasiliencis

9EQ5 の概要

• エントリーDOI: 10.2210/pdb9eq5/pdb
• EMDBエントリー: 19897
• 分子名称: Histidine ammonia-lyase, [(1R)-1-amino-2-phenylethyl]phosphonic acid (2 entities in total)
• 機能のキーワード: phenylalanine catabolism lyase, lyase
• 由来する生物種: Rubinisphaera brasiliensis
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 251645.97

構造登録者

Duhoo, Y.,Buslov, I.,Desmons, S. (登録日: 2024-03-20, 公開日: 2024-08-07, 最終更新日: 2024-10-09)

主引用文献

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

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: 38713131
DOI: 10.1002/anie.202406008

実験手法

ELECTRON MICROSCOPY (2.17 Å)