RT Journal Article
SR Electronic
T1 In vitro biosynthesis of poly-β-1,4-glucan derivatives using a pro-miscuous glycosyltransferase
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.02.14.949545
DO 10.1101/2020.02.14.949545
A1 Gregory S. Bulmer
A1 Ashley P. Mattey
A1 Fabio Parmeggiani
A1 Ryan Williams
A1 Helene Ledru
A1 Andrea Marchesi
A1 Lisa S. Seibt
A1 Peter Both
A1 Kun Huang
A1 M. Carmen Galan
A1 Sabine L. Flitsch
A1 Anthony P. Green
A1 Jolanda M. van Munster
YR 2020
UL http://biorxiv.org/content/early/2020/02/15/2020.02.14.949545.abstract
AB 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.