Phenylalanine Ammonia-Lyase-Catalyzed Deamination of an Acyclic Amino Acid

Enzyme Mechanistic Studies Aided by a Novel Microreactor Filled with Magnetic Nanoparticles

Diána Weiser, Lászlõ Csaba Bencze, Gergely Bánõczi, Ferenc Ender, Rõbert Kiss, Eszter Kõkai, A. Szilágyi, B. Vértessy, O. Farkas, Csaba Paizs, L. Poppe

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Phenylalanine ammonia-lyase (PAL), found in many organisms, catalyzes the deamination of l-phenylalanine (Phe) to (E)-cinnamate by the aid of its MIO prosthetic group. By using PAL immobilized on magnetic nanoparticles and fixed in a microfluidic reactor with an in-line UV detector, we demonstrated that PAL can catalyze ammonia elimination from the acyclic propargylglycine (PG) to yield (E)-pent-2-ene-4-ynoate. This highlights new opportunities to extend MIO enzymes towards acyclic substrates. As PG is acyclic, its deamination cannot involve a Friedel-Crafts-type attack at an aromatic ring. The reversibility of the PAL reaction, demonstrated by the ammonia addition to (E)-pent-2-ene-4-ynoate yielding enantiopure l-PG, contradicts the proposed highly exothermic single-step mechanism. Computations with the QM/MM models of the N-MIO intermediates from l-PG and l-Phe in PAL show similar arrangements within the active site, thus supporting a mechanism via the N-MIO intermediate.

Original languageEnglish
Pages (from-to)2283-2288
Number of pages6
JournalChemBioChem
Volume16
Issue number16
DOIs
Publication statusPublished - Nov 1 2015

Fingerprint

Acyclic Acids
Phenylalanine Ammonia-Lyase
Deamination
Nanoparticles
Amino Acids
Enzymes
Phenylalanine
Ammonia
Cinnamates
Ultraviolet detectors
Microfluidics
Prosthetics
Catalytic Domain
propargylglycine
Substrates

Keywords

  • enzyme catalysis
  • magnetic nanoparticles
  • microreactors
  • phenylalanine ammonia lyase
  • reaction mechanisms

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology

Cite this

Phenylalanine Ammonia-Lyase-Catalyzed Deamination of an Acyclic Amino Acid : Enzyme Mechanistic Studies Aided by a Novel Microreactor Filled with Magnetic Nanoparticles. / Weiser, Diána; Bencze, Lászlõ Csaba; Bánõczi, Gergely; Ender, Ferenc; Kiss, Rõbert; Kõkai, Eszter; Szilágyi, A.; Vértessy, B.; Farkas, O.; Paizs, Csaba; Poppe, L.

In: ChemBioChem, Vol. 16, No. 16, 01.11.2015, p. 2283-2288.

Research output: Contribution to journalArticle

Weiser, Diána ; Bencze, Lászlõ Csaba ; Bánõczi, Gergely ; Ender, Ferenc ; Kiss, Rõbert ; Kõkai, Eszter ; Szilágyi, A. ; Vértessy, B. ; Farkas, O. ; Paizs, Csaba ; Poppe, L. / Phenylalanine Ammonia-Lyase-Catalyzed Deamination of an Acyclic Amino Acid : Enzyme Mechanistic Studies Aided by a Novel Microreactor Filled with Magnetic Nanoparticles. In: ChemBioChem. 2015 ; Vol. 16, No. 16. pp. 2283-2288.
@article{1dfcb1e3cb5b432eaa650ba43465ccb6,
title = "Phenylalanine Ammonia-Lyase-Catalyzed Deamination of an Acyclic Amino Acid: Enzyme Mechanistic Studies Aided by a Novel Microreactor Filled with Magnetic Nanoparticles",
abstract = "Phenylalanine ammonia-lyase (PAL), found in many organisms, catalyzes the deamination of l-phenylalanine (Phe) to (E)-cinnamate by the aid of its MIO prosthetic group. By using PAL immobilized on magnetic nanoparticles and fixed in a microfluidic reactor with an in-line UV detector, we demonstrated that PAL can catalyze ammonia elimination from the acyclic propargylglycine (PG) to yield (E)-pent-2-ene-4-ynoate. This highlights new opportunities to extend MIO enzymes towards acyclic substrates. As PG is acyclic, its deamination cannot involve a Friedel-Crafts-type attack at an aromatic ring. The reversibility of the PAL reaction, demonstrated by the ammonia addition to (E)-pent-2-ene-4-ynoate yielding enantiopure l-PG, contradicts the proposed highly exothermic single-step mechanism. Computations with the QM/MM models of the N-MIO intermediates from l-PG and l-Phe in PAL show similar arrangements within the active site, thus supporting a mechanism via the N-MIO intermediate.",
keywords = "enzyme catalysis, magnetic nanoparticles, microreactors, phenylalanine ammonia lyase, reaction mechanisms",
author = "Di{\'a}na Weiser and Bencze, {L{\'a}szl{\~o} Csaba} and Gergely B{\'a}n{\~o}czi and Ferenc Ender and R{\~o}bert Kiss and Eszter K{\~o}kai and A. Szil{\'a}gyi and B. V{\'e}rtessy and O. Farkas and Csaba Paizs and L. Poppe",
year = "2015",
month = "11",
day = "1",
doi = "10.1002/cbic.201500444",
language = "English",
volume = "16",
pages = "2283--2288",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "Wiley-VCH Verlag",
number = "16",

}

TY - JOUR

T1 - Phenylalanine Ammonia-Lyase-Catalyzed Deamination of an Acyclic Amino Acid

T2 - Enzyme Mechanistic Studies Aided by a Novel Microreactor Filled with Magnetic Nanoparticles

AU - Weiser, Diána

AU - Bencze, Lászlõ Csaba

AU - Bánõczi, Gergely

AU - Ender, Ferenc

AU - Kiss, Rõbert

AU - Kõkai, Eszter

AU - Szilágyi, A.

AU - Vértessy, B.

AU - Farkas, O.

AU - Paizs, Csaba

AU - Poppe, L.

PY - 2015/11/1

Y1 - 2015/11/1

N2 - Phenylalanine ammonia-lyase (PAL), found in many organisms, catalyzes the deamination of l-phenylalanine (Phe) to (E)-cinnamate by the aid of its MIO prosthetic group. By using PAL immobilized on magnetic nanoparticles and fixed in a microfluidic reactor with an in-line UV detector, we demonstrated that PAL can catalyze ammonia elimination from the acyclic propargylglycine (PG) to yield (E)-pent-2-ene-4-ynoate. This highlights new opportunities to extend MIO enzymes towards acyclic substrates. As PG is acyclic, its deamination cannot involve a Friedel-Crafts-type attack at an aromatic ring. The reversibility of the PAL reaction, demonstrated by the ammonia addition to (E)-pent-2-ene-4-ynoate yielding enantiopure l-PG, contradicts the proposed highly exothermic single-step mechanism. Computations with the QM/MM models of the N-MIO intermediates from l-PG and l-Phe in PAL show similar arrangements within the active site, thus supporting a mechanism via the N-MIO intermediate.

AB - Phenylalanine ammonia-lyase (PAL), found in many organisms, catalyzes the deamination of l-phenylalanine (Phe) to (E)-cinnamate by the aid of its MIO prosthetic group. By using PAL immobilized on magnetic nanoparticles and fixed in a microfluidic reactor with an in-line UV detector, we demonstrated that PAL can catalyze ammonia elimination from the acyclic propargylglycine (PG) to yield (E)-pent-2-ene-4-ynoate. This highlights new opportunities to extend MIO enzymes towards acyclic substrates. As PG is acyclic, its deamination cannot involve a Friedel-Crafts-type attack at an aromatic ring. The reversibility of the PAL reaction, demonstrated by the ammonia addition to (E)-pent-2-ene-4-ynoate yielding enantiopure l-PG, contradicts the proposed highly exothermic single-step mechanism. Computations with the QM/MM models of the N-MIO intermediates from l-PG and l-Phe in PAL show similar arrangements within the active site, thus supporting a mechanism via the N-MIO intermediate.

KW - enzyme catalysis

KW - magnetic nanoparticles

KW - microreactors

KW - phenylalanine ammonia lyase

KW - reaction mechanisms

UR - http://www.scopus.com/inward/record.url?scp=84945183335&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84945183335&partnerID=8YFLogxK

U2 - 10.1002/cbic.201500444

DO - 10.1002/cbic.201500444

M3 - Article

VL - 16

SP - 2283

EP - 2288

JO - ChemBioChem

JF - ChemBioChem

SN - 1439-4227

IS - 16

ER -