Tailored Mutants of Phenylalanine Ammonia-Lyase from Petroselinum crispum for the Synthesis of Bulky l- and d-Arylalanines

Alina Filip, Emma Z.A. Nagy, Souad D. Tork, Gergely Bánóczi, Monica I. Toşa, Florin D. Irimie, László Poppe, Csaba Paizs, László C. Bencze

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Tailored mutants of phenylalanine ammonia-lyase from Petroselinum crispum (PcPAL) were created and tested in ammonia elimination from various sterically demanding, non-natural analogues of phenylalanine and in ammonia addition reactions into the corresponding (E)-arylacrylates. The wild-type PcPAL was inert or exhibited quite poor conversions in both reactions with all members of the substrate panel. Appropriate single mutations of residue F137 and the highly conserved residue I460 resulted in PcPAL variants that were active in ammonia elimination but still had a poor activity in ammonia addition onto bulky substrates. However, combined mutations that involve I460 besides the well-studied F137 led to mutants that exhibited activity in ammonia addition as well. The synergistic multiple mutations resulted in substantial substrate scope extension of PcPAL and opened up new biocatalytic routes for the synthesis of both enantiomers of valuable phenylalanine analogues, such as (4-methoxyphenyl)-, (napthalen-2-yl)-, ([1,1'-biphenyl]-4-yl)-, (4'-fluoro-[1,1'-biphenyl]-4-yl)-, and (5-phenylthiophene-2-yl)alanines.

Original languageEnglish
JournalChemCatChem
DOIs
Publication statusAccepted/In press - Jan 1 2018

Fingerprint

Phenylalanine Ammonia-Lyase
phenylalanine
Ammonia
ammonia
synthesis
mutations
Phenylalanine
Substrates
elimination
Addition reactions
Enantiomers
analogs
Alanine
enantiomers
alanine
routes

Keywords

  • Amino acids
  • Biocatalysis
  • Phenylalanine ammonia-lyase
  • Protein engineering
  • Substrate scope extension

ASJC Scopus subject areas

  • Catalysis
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

Cite this

Filip, A., Nagy, E. Z. A., Tork, S. D., Bánóczi, G., Toşa, M. I., Irimie, F. D., ... Bencze, L. C. (Accepted/In press). Tailored Mutants of Phenylalanine Ammonia-Lyase from Petroselinum crispum for the Synthesis of Bulky l- and d-Arylalanines. ChemCatChem. https://doi.org/10.1002/cctc.201800258

Tailored Mutants of Phenylalanine Ammonia-Lyase from Petroselinum crispum for the Synthesis of Bulky l- and d-Arylalanines. / Filip, Alina; Nagy, Emma Z.A.; Tork, Souad D.; Bánóczi, Gergely; Toşa, Monica I.; Irimie, Florin D.; Poppe, László; Paizs, Csaba; Bencze, László C.

In: ChemCatChem, 01.01.2018.

Research output: Contribution to journalArticle

Filip, Alina ; Nagy, Emma Z.A. ; Tork, Souad D. ; Bánóczi, Gergely ; Toşa, Monica I. ; Irimie, Florin D. ; Poppe, László ; Paizs, Csaba ; Bencze, László C. / Tailored Mutants of Phenylalanine Ammonia-Lyase from Petroselinum crispum for the Synthesis of Bulky l- and d-Arylalanines. In: ChemCatChem. 2018.
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