1. 2D and 3D maximum-quantum NMR and diffusion spectroscopy for the characterization of enzymatic reaction mixtures
- Author
-
Elena Piersanti, Claudio Righetti, David Ribeaucourt, A. Jalila Simaan, Yasmina Mekmouche, Mickael Lafond, Jean-Guy Berrin, Thierry Tron, Mehdi Yemloul, Institut des Sciences Moléculaires de Marseille (ISM2), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Biodiversité et Biotechnologie Fongiques (BBF), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), V. Mane Fils, 620 route de Grasse, 06620 Le Bar sur Loup, France., and ANR-17-CE07-0047,FUNTASTIC,Nouvelles oxydases à radical cuivre fongiques utilisées comme biocatalyseurs pour la valorisation de la biomasse végétale et d'alcools(2017)
- Subjects
Magnetic Resonance Spectroscopy ,[CHIM.ANAL]Chemical Sciences/Analytical chemistry ,Electrochemistry ,Environmental Chemistry ,[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM] ,Biochemistry ,Magnetic Resonance Imaging ,Spectroscopy ,Analytical Chemistry - Abstract
International audience; 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.
- Published
- 2022