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.

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.

Citation

Keenan, T., Hatton, N. E., Porter, J., Vendeville, J., Wheatley, D. E., Ghirardello, M., …Fascione, M. A. (in press). Reverse thiophosphorylase activity of a glycoside phosphorylase in the synthesis of an unnatural Manβ1,4GlcNAc library †. Chemical Science, 14(42), 11638-11646. https://doi.org/10.1039/d3sc04169g

• 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