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Enzymatic synthesis of some (15)N-labelled L-amino acids.

Isotopes in environmental and health studies (2010-06-29)
Maria Chiriac, Iulia Lupan, Flavia Popa, Nicolae Palibroda, Octavian Popescu
ZUSAMMENFASSUNG

Our group has developed a stereospecific enzymatic method, which is very efficient for the in vitro synthesis of l-[(15)N]serine, l-[(15)N]methionine and l-[(15)N]glutamic acid. These amino acids were prepared from the corresponding alpha -ketoacids in the suitable enzymatic systems. The bacterial NAD-dependent amino acid dehydrogenases alanin dehydrogenase, leucin dehydrogenase and glutamate dehydrogenase were used as catalysts. Glucose dehydrogenase was used for the regeneration of NADH and (15)NH(4)Cl as isotopically labelled material at 99 at.% (15)N. All reactions are inexpensive and easy to perform on a synthetically useful scale (1-10g) giving high yields of l-amino acids. The (15)N isotope content was determined by mass spectrometry.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
L-Alanin-Dehydrogenase aus Bacillus subtilis, buffered aqueous glycerol solution, ~30 units/mg protein (Lowry)
Sigma-Aldrich
Alanin-Dehydrogenase, rekombinant, recombinant, expressed in E. coli, ≥15 U/mg
Sigma-Aldrich
L-Leucin-Dehydrogenase aus Bacillus cereus, lyophilized powder, ≥60 units/mg protein
Sigma-Aldrich
L-Alanin-Dehydrogenase aus Bacillus subtilis, ammonium sulfate suspension, ≥20 units/mg protein (Lowry)