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Updated: Jul 14, 2026
Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
Published on: August 8, 2016
Selective detection of sugar phosphates by capillary electrophoresis/mass spectrometry and its application to an
Joseph P M Hui1, Jie Yang, Jon S Thorson
1MS Metabolomics Group, NRC-Institute for Marine Biosciences, 1411 Oxford Street, Halifax, NS B3H 3Z1, Canada.
Abstract:
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
