title: "On-the-fly" kinetics of enzymatic racemization using deuterium NMR in DNA-based chiral oriented media.
abstract: We report the in situ and real-time monitoring of the interconversion of L- and D-alanine-d3 by alanine racemase from Bacillus stearothermophilus directly observed by (2)H NMR spectroscopy in anisotropic phase. The enantiomers are distinguished by the difference of their (2)H quadrupolar splittings in a chiral liquid crystal containing 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.
authors.affiliation: Institut Curie, Centre de Recherche, 91405 Orsay Cedex, France. chanhuot.monique@gmail.com
authors.forename: Monique
authors.initials: M
authors.lastname: Chan-Huot
authors.forename: Philippe
authors.initials: P
authors.lastname: Lesot
authors.forename: Philippe
authors.initials: P
authors.lastname: Pelupessy
authors.forename: Luminita
authors.initials: L
authors.lastname: Duma
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Bodenhausen
authors.forename: Patricia
authors.initials: P
authors.lastname: Duchambon
authors.forename: Michael D
authors.initials: MD
authors.lastname: Toney
authors.forename: U Venkateswara
authors.initials: UV
authors.lastname: Reddy
authors.forename: N
authors.initials: N
authors.lastname: Suryaprakash
mesh_terms: D000409:Alanine / Q000737:chemistry / Q000378:metabolism; D000411:Alanine Racemase / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D003903:Deuterium / Q000737:chemistry*; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D007700:Kinetics; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D013237:Stereoisomerism
chemical_list: D004247:DNA; D003903:Deuterium; D000411:Alanine Racemase; D000409:Alanine