Isotopic effects in mechanistic studies of biotransformations of fluorine derivatives of L-alanine catalysed by L-alanine dehydrogenase

Jolanta Szymańska-Majchrzak; Katarzyna Pałka; Marianna Kańska

doi:10.1016/j.apradiso.2017.02.003

Isotopic effects in mechanistic studies of biotransformations of fluorine derivatives of L-alanine catalysed by L-alanine dehydrogenase

Appl Radiat Isot. 2017 May:123:21-25. doi: 10.1016/j.apradiso.2017.02.003. Epub 2017 Feb 12.

Authors

Jolanta Szymańska-Majchrzak¹, Katarzyna Pałka², Marianna Kańska³

Affiliations

¹ Department of Biochemistry, 2nd Faculty of Medicine, Medical University of Warsaw, 101 Żwirki i Wigury Ave., 02-089 Warsaw, Poland. Electronic address: jolanta.szymanska@wum.edu.pl.
² Department of Chemistry, University of Warsaw, 1 Pasteur Str., 02-093 Warsaw, Poland.
³ Department of Biochemistry, 2nd Faculty of Medicine, Medical University of Warsaw, 101 Żwirki i Wigury Ave., 02-089 Warsaw, Poland.

PMID: 28209535
DOI: 10.1016/j.apradiso.2017.02.003

Abstract

Synthesis of 3-fluoro-[2-²H]-L-alanine (3-F-[²H]-L-Ala) in reductive amination of 3-fluoropyruvic acid catalysed by L-alanine dehydrogenase (AlaDH) was described. Fluorine derivative was used to study oxidative deamination catalysed by AlaDH applied kinetic (for 3-F-L-Ala in H₂O - KIE's on V_max: 1.1; on V_max/K_M: 1.2; for 3-F-L-Ala in ²H₂O - on V_max: 1.4; on V_max/K_M: 2.1) and solvent isotope effect methods (for 3-F-L-Ala - SIE's on V_max: 1.0; on V_max/K_M: 0.87; for 3-F-[2-²H]-L-Ala - on V_max: 1.4; on V_max/K_M: 1.5). Studies explain some details of reaction mechanism.

Keywords: Deuterium; Fluorine; Isotope effect; L-alanine; L-alanine dehydrogenase; UV–VIS spectrophotometry.

MeSH terms

Alanine / analogs & derivatives*
Alanine / chemistry
Alanine / metabolism
Alanine Dehydrogenase / metabolism*
Biocatalysis
Biotransformation
Deamination
Deuterium / chemistry
Kinetics
Models, Chemical
Oxidation-Reduction
Solvents

Substances

Solvents
3-fluoroalanine
Deuterium
Alanine Dehydrogenase
Alanine