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Stereo-specificity for pro-(R) hydrogen of NAD(P)H during enzyme-catalyzed hydride transfer to CL-20

Par Conseil national de recherches du Canada

DOITrouver le DOI : https://doi.org/10.1016/j.bbrc.2005.09.166
AuteurRechercher : 1; Rechercher : 1; Rechercher : 1
Affiliation
  1. Conseil national de recherches du Canada. Institut de recherche en biotechnologie du CNRC
FormatTexte, Article
Résumé
A dehydrogenase from Clostridium sp. EDB2 and a diaphorase from Clostridium kluyveri were reacted with CL-20 to gain insights into the enzyme-catalyzed hydride transfer to CL-20, and the enzyme's stereo-specificity for either pro-R or pro-S hydrogens of NAD(P)H. Both enzymes biotransformed CL-20 at rates of 18.5 and 24 nmol/h/mg protein, using NADH and NADPH as hydride-source, respectively, to produce a N-denitrohydrogenated product with a molecular weight of 393 Da. In enzyme kinetics studies using reduced deuterated pyridine nucleotides, we found a kinetic deuterium isotopic effect of 2-fold on CL-20 biotransformation rate using dehydrogenase enzyme against (R)NADD as a hydride-source compared to either (S)NADD or NADH. Whereas, in case of diaphorase, the kinetic deuterium isotopic effect of about 1.5-fold was observed on CL-20 biotransformation rate using (R)NADPD as hydride-source. In a comparative study with LC-MS, using deuterated and non-deuterated NAD(P)H, we found a positive mass-shift of 1 Da in the N-denitrohydrogenated product suggesting the involvement of a deuteride (D-) transfer from NAD(P)D. The present study thus revealed that both dehydrogenase and diaphorase enzymes from the two Clostridium species catalyzed a hydride transfer to CL-20 and showed stereo-specificity for pro-R hydrogen of NAD(P)H.
Date de publication
Dans
  • Biochemical and Biophysical Research Communications 337, nº 4 : 1080–1083.
Langueanglais
Numéro du CNRC47245
Numéro NPARC3540086
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Identificateur de l’enregistrement59a735d0-38a3-4713-944b-777d3b26c9f0
Enregistrement créé2009-03-01
Enregistrement modifié2020-04-07
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