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

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DOIResolve DOI: http://doi.org/10.1002/cbic.200700116
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TypeArticle
Journal titleChemoBioChem
Volume8
Issue10
Pages11801188; # of pages: 9
Subjectanalytical methods; bioorganic chemistry; electrophoresis; glycorandomization; mass spectrometry; sugar phosphates
AbstractA 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.
Publication date
PublisherWiley
Copyright noticeCopyright © 2007 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
LanguageEnglish
AffiliationNRC Institute for Marine Biosciences; National Research Council Canada; Aquatic and Crop Resource Development
Peer reviewedYes
NRC number42705
1242
NPARC number3538056
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Record identifier80a469eb-9cbb-43ac-b86d-1307b1370362
Record created2009-03-01
Record modified2016-05-09
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