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Reverse thiophosphorylase activity of a glycoside phosphorylase in the synthesis of an unnatural Manβ1,4GlcNAc library †

Keenan, Tessa; Hatton, Natasha E.; Porter, Jack; Vendeville, Jean-Baptiste; Wheatley, David E.; Ghirardello, Mattia; Wahart, Alice. J. C.; Ahmadipour, Sanaz; Walton, Julia; Galan, M. Carmen; Linclau, Bruno; Miller, Gavin J.; Fascione, Martin A.

Authors

Tessa Keenan

Natasha E. Hatton

Jean-Baptiste Vendeville

David E. Wheatley

Mattia Ghirardello

Alice. J. C. Wahart

Julia Walton

M. Carmen Galan

Bruno Linclau

Martin A. Fascione



Abstract

β-Mannosides are ubiquitous in nature, with diverse roles in many biological processes. Notably, Manβ1,4GlcNAc a constituent of the core N-glycan in eukaryotes was recently identified as an immune activator, highlighting its potential for use in immunotherapy. Despite their biological significance, the synthesis of β-mannosidic linkages remains one of the major challenges in glycoscience. Here we present a chemoenzymatic strategy that affords a series of novel unnatural Manβ1,4GlcNAc analogues using the β-1,4-d-mannosyl-N-acetyl-d-glucosamine phosphorylase, BT1033. We show that the presence of fluorine in the GlcNAc acceptor facilitates the formation of longer β-mannan-like glycans. We also pioneer a “reverse thiophosphorylase” enzymatic activity, favouring the synthesis of longer glycans by catalysing the formation of a phosphorolysis-stable thioglycoside linkage, an approach that may be generally applicable to other phosphorylases.

Journal Article Type Article
Acceptance Date Sep 28, 2023
Online Publication Date Sep 29, 2023
Deposit Date Oct 9, 2023
Publicly Available Date Oct 9, 2023
Journal Chemical Science
Print ISSN 2041-6520
Electronic ISSN 2041-6539
Publisher Royal Society of Chemistry
Peer Reviewed Peer Reviewed
Volume 14
Issue 42
Pages 11638-11646
DOI https://doi.org/10.1039/d3sc04169g