In vitro biosynthesis of poly-β-1,4-glucan derivatives using a pro-miscuous glycosyltransferase
| Autor: | van Munster Jm, Fabio Parmeggiani, Ashley P. Mattey, Peter Both, Helene Ledru, Gregory S Bulmer, Rachel Williams, Lisa S. Seibt, Kun Huang, Sabine L. Flitsch, Anthony P. Green, Galan Mc, Andrea Marchesi |
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| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Glycan biology 010402 general chemistry Polysaccharide 01 natural sciences 0104 chemical sciences Cell wall 03 medical and health sciences chemistry.chemical_compound chemistry Biochemistry Biocatalysis Glycosyltransferase biology.protein Nanobiotechnology Cellulose 030304 developmental biology Glucan |
| DOI: | 10.1101/2020.02.14.949545 |
| Popis: | The β-1,4-glucose linkage of cellulose is the most abundant polymeric linkage on earth and as such is of considerable interest in biology and biotechnology. It remains challenging to synthesize this linkage in vitro due to a lack of suitable biocatalysts; the natural cellulose biosynthetic machinery is a membrane-associated complex with processive activity that cannot be easily manipulated to synthesize tailor-made oligosaccharides and their derivatives. Here we identify a promiscuous activity of a soluble recombinant biocatalyst, Neisseria meningitidis glycosyltransferase LgtB, suitable for the polymerization of glucose from UDP-glucose via the generation of β-1,4-glycosidic linkages. We employed LgtB to synthesize natural and derivatized cello-oligosaccharides and we demonstrate how LgtB can be incorporated in biocatalytic cascades and chemo-enzymatic strategies to synthesize cello-oligosaccharides with tailored functionalities. We also show how the resulting glycan structures can be applied as chemical probes to report on activity and selectivity of plant cell wall degrading enzymes, including lytic polysaccharide monooxygenases. We anticipate that this biocatalytic approach to derivatized cello-oligosaccharides via glucose polymerization will open up new applications in biology and nanobiotechnology. |
| Databáze: | OpenAIRE |
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