Reverse thiophosphorylase activity of a glycoside phosphorylase in the synthesis of an unnatural Manβ1,4GlcNAc library
Reverse thiophosphorylase activity of a glycoside phosphorylase in the synthesis of an unnatural Manβ1,4GlcNAc library
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.
11638-11646
Keenan, Tessa
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Hatton, Natasha E.
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Porter, Jack
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Vendeville, Jean-Baptiste
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Wheatley, David
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Ghirardello, Mattia
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Wahart, Alice. J.C.
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Ahmadipour, Sanaz
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Walton, Julia
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Galan, M. Carmen
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Linclau, Bruno
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Miller, Gavin J.
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Fascione, Martin A.
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29 September 2023
Keenan, Tessa
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Hatton, Natasha E.
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Porter, Jack
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Vendeville, Jean-Baptiste
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Wheatley, David
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Ghirardello, Mattia
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Wahart, Alice. J.C.
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Ahmadipour, Sanaz
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Walton, Julia
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Galan, M. Carmen
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Linclau, Bruno
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Miller, Gavin J.
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Fascione, Martin A.
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Keenan, Tessa, Hatton, Natasha E., Porter, Jack, Vendeville, Jean-Baptiste, Wheatley, David, Ghirardello, Mattia, Wahart, Alice. J.C., Ahmadipour, Sanaz, Walton, Julia, Galan, M. Carmen, Linclau, Bruno, Miller, Gavin J. and Fascione, Martin A.
(2023)
Reverse thiophosphorylase activity of a glycoside phosphorylase in the synthesis of an unnatural Manβ1,4GlcNAc library.
Chemical Science, 14 (42), .
(doi:10.1039/d3sc04169g).
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.
Text
d3sc04169g
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Accepted/In Press date: 28 August 2023
e-pub ahead of print date: 29 September 2023
Published date: 29 September 2023
Additional Information:
Funding Information:
MAF thanks UKRI (EP/X023680/1) for project grant funding. UK Research and Innovation (UKRI, Future Leaders Fellow-ship, MR/T019522/1) and the Engineering and Physical Sciences Research Council (EP/P000762/1) are thanked for project grant funding to GJM. Keele University are thanked for PhD studentship funding to AW and JP. BL thanks EPSRC core capability funding EP/K039466/1. MCG thanks Cancer Research UK (grant number C30758/A2979) and European Research Council (ERC-COG GLYCOTOOLS 648239). MAF, BL and MCG are grateful to the Industrial Biotechnology Catalyst (Innovate UK, BBSRC, EPSRC) to support the translation, development and commercialization of innovative Industrial Biotechnology processes: BB/M028941/1, BB/M02847X/1, BB/M028976/1. We thank Dr Ed Bergstrom and The York Center of Excellence in Mass Spectrometry was created thanks to a major capital investment through Science City York, supported by Yorkshire Forward with funds from the Northern Way Initiative, and subsequent support from EPSRC (EP/K039660/1; EP/M028127/1). We thank Dr Alex Heyam for discussions around the IPAP NMR experiments.
Publisher Copyright:
© 2023 The Royal Society of Chemistry
Identifiers
Local EPrints ID: 485395
URI: http://eprints.soton.ac.uk/id/eprint/485395
ISSN: 1478-6524
PURE UUID: d8e344e6-a2f2-4573-a930-430037aeba85
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Date deposited: 05 Dec 2023 17:57
Last modified: 18 Mar 2024 02:52
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Contributors
Author:
Tessa Keenan
Author:
Natasha E. Hatton
Author:
Jack Porter
Author:
Jean-Baptiste Vendeville
Author:
David Wheatley
Author:
Mattia Ghirardello
Author:
Alice. J.C. Wahart
Author:
Sanaz Ahmadipour
Author:
Julia Walton
Author:
M. Carmen Galan
Author:
Gavin J. Miller
Author:
Martin A. Fascione
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