Preparative and Kinetic Analysis of β-1,4- and β-1,3-Glucan Phosphorylases Informs Access to Human Milk Oligosaccharide Fragments and Analogues Thereof

Singh, Ravindra Pal, Pergolizzi, Giulia, Nepogodiev, Sergey A., de Andrade, Peterson, Kuhaudomlarp, Sakonwan and Field, Robert A. (2020) Preparative and Kinetic Analysis of β-1,4- and β-1,3-Glucan Phosphorylases Informs Access to Human Milk Oligosaccharide Fragments and Analogues Thereof. ChemBioChem, 21 (7). pp. 1043-1049. ISSN 1439-4227

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Abstract

The enzymatic synthesis of oligosaccharides depends on the availability of suitable enzymes, which remains a limitation. Without recourse to enzyme engineering or evolution approaches, herein we demonstrate the ability of wild-type cellodextrin phosphorylase (CDP: β-1,4-glucan linkage-dependent) and laminaridextrin phosphorylase (Pro_7066: β-1,3-glucan linkage-dependent) to tolerate a number of sugar-1- phosphate substrates, albeit with reduced kinetic efficiency. In spite of catalytic efficiencies of <1 % of the natural reactions, we demonstrate the utility of given phosphorylase–sugar phosphate pairs to access new-to-nature fragments of human milk oligosaccharides, or analogues thereof, in multi-milligram quantities.

Item Type: Article
Additional Information: Funding Information: These studies were supported by the UK BBSRC Institute Strategic Program on Molecules from Nature—Products and Pathways [BBS/E/J/000PR9790] and the John Innes Foundation; the BBSRC and ESRC–Newton Fund [BB/N005082/1], the Department for International Development, and the Indian Department of Biotechnology under the Newton Fund Global Research Partnership in Aquaculture programme; and the InnovateUK IBCatalyst [BB/M02903411 and EP/N033167/10]. We thank Gerhard Saalbach for assistance with the mass spectrometry. Publisher Copyright: © 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.
Uncontrolled Keywords: enzymatic synthesis,glycans,oligosaccharides,phosphorylases,biochemistry,molecular medicine,molecular biology,organic chemistry ,/dk/atira/pure/subjectarea/asjc/1300/1303
Faculty \ School:
Faculty of Science > School of Chemistry, Pharmacy and Pharmacology
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Depositing User: LivePure Connector
Date Deposited: 02 Sep 2024 15:30
Last Modified: 28 Jan 2025 23:26
URI: https://ueaeprints.uea.ac.uk/id/eprint/96436
DOI:

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