Direct asymmetric synthesis of β-branched aromatic α-amino acids using engineered phenylalanine ammonia lyases.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
26 Sep 2024
26 Sep 2024
Historique:
received:
14
03
2024
accepted:
16
09
2024
medline:
27
9
2024
pubmed:
27
9
2024
entrez:
26
9
2024
Statut:
epublish
Résumé
β-Branched aromatic α-amino acids are valuable building blocks in natural products and pharmaceutically active compounds. However, their chemical or enzymatic synthesis is challenging due to the presence of two stereocenters. We design phenylalanine ammonia lyases (PAL) variants for the direct asymmetric synthesis of β-branched aromatic α-amino acids. Based on extensive computational analyses, we unravel the enigma behind PAL's inability to accept β-methyl cinnamic acid (β-MeCA) as substrate and achieve the synthesis of the corresponding amino acids of β-MeCA and analogs using a double (PcPAL-L256V-I460V) and a triple mutant (PcPAL-F137V-L256V-I460V). The reactions are scaled-up using an optimized E. coli based whole-cell biotransformation system to produce ten β-branched phenylalanine analogs with high diastereoselectivity (dr > 20:1) and enantioselectivity (ee > 99.5%) in yields ranging from 41-71%. Moreover, we decipher the mechanism of PcPAL-L256V-I460V for the acceptance of β-MeCA and converting it with excellent stereoselectivity by computational simulations. Thus, this study offers an efficient method for synthesizing β-branched aromatic α-amino acids.
Identifiants
pubmed: 39327443
doi: 10.1038/s41467-024-52613-x
pii: 10.1038/s41467-024-52613-x
doi:
Substances chimiques
Phenylalanine Ammonia-Lyase
EC 4.3.1.24
Amino Acids, Aromatic
0
Phenylalanine
47E5O17Y3R
Cinnamates
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
8264Subventions
Organisme : EC | Horizon 2020 Framework Programme (EU Framework Programme for Research and Innovation H2020)
ID : 814650
Organisme : Huazhong University of Science and Technology (HUST)
ID : 3034514105
Informations de copyright
© 2024. The Author(s).
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