Hyperpolarized NMR Analysis of Enzymatic Reaction: Extension of Observable Reaction Time by Deuterium Isotope Labeling

Abstract

Affording several orders of signal enhancement compared to conventional nuclear magnetic resonance (NMR), dynamic nuclear polarization (DNP) enables direct observation of the most common C-13 nuclei in organic chemistry and biochemistry at low concentrations. These nuclei have poor sensitivity due to low natural abundance and a low gyromagnetic ratio. Herein, we report an application of this technique to a multi-step enzymatic reaction. The main aim of this study was to demontrate the effect of deuteration on specific spins of interest for the real-time trypsin hydrolysis reaction of synthesized N-alpha-benzoyl-L-arginine methyl-d(3) ester (BAME-d(3)). Kinetic parameter and fit accuracy were analyzed in order to determine the influence of deuteration.