Application of 2H NMR to the study of natural site-specific hydrogen isotope transfer among substrate, medium, and glycerol in glucose fermentation with yeast.
2H NMR is a very useful tool in isotope tracing studies. This technique was applied to a quantitative study of a site-specific deuterium affiliation among the substrate, the medium, and a product (glycerol), in glucose fermentation with yeast. The quality of the results depends on the quantitative 2H NMR analysis of glycerol. After comparing several potential analysis probe molecules, the derivative of glycerol, 2,2-dimethyl-1,3-dioxolane-4-methanol, was chosen as the most advantageous. Using this probe in a set of isotope-labeling experiments, we describe how a complete quantitative site-specific hydrogen isotope transfer model, which connects the site-specific isotopic ratios of the substrate, the medium, and the products, can be established. This model can provide information on complex hydrogen transfer mechanisms during biochemical reactions and can be useful for the prediction of site-specific hydrogen isotopic ratios at natural abundance of the products, based on that of the substrate or reactants and the medium.
Laboratoire d'Analyse Isotopique et Electrochimique de Métabolismes, CNRS UMR 6006, Université de Nantes, 2, rue de la Houssinière, 44322 Nantes, France.
Sébastien
S
Pionnier
Ben-Li
BL
Zhang
D003470:Culture Media; D003903:Deuterium / Q000737:chemistry; D005285:Fermentation; D005947:Glucose / Q000378:metabolism*; D005990:Glycerol / Q000378:metabolism*; D006859:Hydrogen / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D003313:Zea mays / Q000737:chemistry
D003470:Culture Media; D006859:Hydrogen; D003903:Deuterium; D005947:Glucose; D005990:Glycerol