On-the-fly' Kinetics of Enzymatic Racemization using Deuterium NMR in DNA-based Chiral Oriented Media.

Monique Chan-Huot (1, 2) , Philippe Georges Julien Lesot (3, 4) , Philippe Pelupessy (5, 6) , Luminita Duma (5, 6) , Geoffrey Bodenhausen (5, 7) , Patricia Duchambon (8, 1) , Michael D Toney (9) , U Venkateswara Reddy (10) , Nagarajarao Suryaprakash (10)

1 Institut Curie [Paris]
2 INSERM - Institut National de la Santé et de la Recherche Médicale
3 ICMMO - Institut de Chimie Moléculaire et des Matériaux d'Orsay
4 CNRS - Centre National de la Recherche Scientifique
5 UPMC - Université Pierre et Marie Curie - Paris 6
6 ENS-PSL - École normale supérieure - Paris
7 EPFL - Ecole Polytechnique Fédérale de Lausanne
8 Imagerie intégrative de la molécule à l'organisme
9 UC Davis - University of California [Davis]
10 IISc Bangalore - Indian Institute of Science [Bangalore]

We report the first in situ and real-time monitoring of the interconversion of L- and D-alanine-d3 by alanine racemase from Bacillus Stearothermophilus directly observed by 2H-NMR spectroscopy in anisotropic phase. The enantiomers are distinguished by the difference of their 2H quadrupolar splittings in a chiral liquid-crystalline environment made of short DNA fragments. The proof-of-principle, the reliability and the robustness of this new method is demonstrated by the determination of the turnover rates of the enzyme using the Michaelis-Menten model.

Mots clés: Polynucleotide liquid crystal; Deuterium; Racemase; DNA; NMR; Turnover rate; Michaelis-Menten model

Domaines: Chimie organique

Monique Chan-Huot, Philippe Georges Julien Lesot, Philippe Pelupessy, Luminita Duma, Geoffrey Bodenhausen, et al.. On-the-fly' Kinetics of Enzymatic Racemization using Deuterium NMR in DNA-based Chiral Oriented Media.. Analytical Chemistry, 2013, 85 (9), pp.4694-4697. ⟨10.1021/ac4004002⟩. ⟨hal-00807811⟩