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.-B., Wheatley, D. E., Ghirardello, M., Wahart, A. J. C., Ahmadipour, S., Walton, J., Galan, M. C., Linclau, B., Miller, G. J., & 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
• Public URL: https://keele-repository.worktribe.com/output/594595