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Selective detection of sugar phosphates by capillary electrophoresis/mass spectrometry and its application to an engineered E. coli host.

Par Conseil national de recherches du Canada

DOITrouver le DOI : https://doi.org/10.1002/cbic.200700116
AuteurRechercher : 1; Rechercher : ; Rechercher : ; Rechercher : 2
Affiliation
  1. Conseil national de recherches du Canada. Développement des cultures et des ressources aquatiques
  2. Conseil national de recherches du Canada. Institut des biosciences marines du CNRC
FormatTexte, Article
Sujetanalytical methods; bioorganic chemistry; electrophoresis; glycorandomization; mass spectrometry; sugar phosphates
Résumé
A highly selective method employing capillary electrophoresis and electrospray mass spectrometry (CE-ESMS) with precursor ion scanning for fragment ions characteristic of phosphate-linked sugars was developed for the determination of unnatural sugar phosphates generated in vivo, as part of a natural product glycorandomization study. Cell lysates from an engineered E. coli host were probed for natural and unnatural sugar phosphates resulting from in vivo galactokinase (GalK) bioconversions, and tandem mass spectrometry experiments were performed to confirm the identities of the sugar phosphates. Among the 22 cell lysates that were studied, 13 were found to contain the expected natural and unnatural sugar phosphates. This was in agreement with the GalK in vitro conversion yields, in which an in vitro yield of 15 % coincided with a lack of observable in vivo bioconversion. In addition, the CE-ESMS and precursor ion scanning method was capable of separating sugar phosphate regioisomers such as hexose-6-phosphate and hexose-1-phosphate.
Date de publication
Conditions d’utilisation
  • Copyright © 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Dans
  • ChemoBioChem 8, nº 10 (9 juillet 2007) : 1180–1188.
Langueanglais
Publications évaluées par des pairsOui
Numéro du CNRCNRCC 42705
1242
Numéro NPARC3538056
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Identificateur de l’enregistrement80a469eb-9cbb-43ac-b86d-1307b1370362
Enregistrement créé2009-03-01
Enregistrement modifié2020-05-10
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