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Increasing field strength versus advanced isotope labeling for NMR‐based fluxomics
Author(s) -
Dinclaux Mickael,
Cahoreau Edern,
Millard Pierre,
Létisse Fabien,
Lippens Guy
Publication year - 2020
Publication title -
magnetic resonance in chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.483
H-Index - 72
eISSN - 1097-458X
pISSN - 0749-1581
DOI - 10.1002/mrc.4988
Subject(s) - chemistry , heteronuclear molecule , isoleucine , deuterium , valine , stable isotope labeling by amino acids in cell culture , isotope , isotopic labeling , nuclear magnetic resonance spectroscopy , field (mathematics) , biosynthesis , leucine , nuclear magnetic resonance , amino acid , stereochemistry , organic chemistry , biochemistry , nuclear physics , physics , gene , enzyme , mathematics , pure mathematics , proteomics
Nuclear magnetic resonance (NMR)‐based fluxomics seeks to measure the incorporation of isotope labels in selected metabolites to follow kinetically the synthesis of the latter. It can however equally be used to understand the biosynthetic origin of the same metabolites. We investigate here different NMR approaches to optimize such experiments in terms of resolution and time requirement. Using the isoleucine biosynthesis as an example, we explore the use of different field strengths ranging from 500 MHz to 1.1 GHz. Because of the different field dependence of chemical shift and heteronuclear J couplings, the spectra change at different field strengths. We equally explore the approach to silence the leucine/valine methyl signals through the use of a suitable deuterated precursor, thereby allowing selective observation of the Ile 13 C labeling pattern. Combining both approaches, we arrive at an efficient procedure for the NMR‐based exploration of Ile biosynthesis.