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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
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

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