2D and 3D Maximum-Quantum NMR and diffusion spectroscopy for the characterization of enzymatic reaction mixtures - Aix-Marseille Université Access content directly
Journal Articles Analyst Year : 2022

2D and 3D Maximum-Quantum NMR and diffusion spectroscopy for the characterization of enzymatic reaction mixtures

Abstract

1D 1H NMR spectroscopy has been widely used to monitor enzymatic activity by recording the evolution of the spectra of substrates and/or products thanks to the linear response of NMR. For complex systems involving the coexistence of multiple compounds (substrate, final product and various intermediates), the identification and quantification can be a more arduous task. Here, we present a simple analytical method for a rapid characterization of reaction mixtures involving enzymatic complexes using Maximum Quantum (MaxQ) NMR, accelerated with the Non-Uniform Sampling (NUS) acquisition procedure. Specifically, this approach enables, in a first analytical step, to count the molecules present in the samples. We also show, using two different enzymatic systems, that the implementation of these pulse sequences implies precautions related to the short relaxation times due to the presence of metallo-enzymes or paramagnetic catalysts. Finally, the combination of MaxQ and diffusion experiments, which leads to a 3D chart, greatly improves the resolution and offers an extreme simplification of the spectra while giving valuable indications on the affinity of the enzymes to the different compounds present in the reaction mixture.
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Dates and versions

hal-03652618 , version 1 (29-04-2022)

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Elena Piersanti, Claudio Righetti, David Ribeaucourt, A. Jalila Simaan, Yasmina Mekmouche, et al.. 2D and 3D Maximum-Quantum NMR and diffusion spectroscopy for the characterization of enzymatic reaction mixtures. Analyst, 2022, 147 (11), pp.2515-2522. ⟨10.1039/D2AN00200K⟩. ⟨hal-03652618⟩
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