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Enzymatic Synthesis of Xylan Microparticles with Tunable Morphologies

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posted on 05.04.2022, 17:49 by Peter J. Smith, Thomas M. Curry, Jeong-Yeh Yang, William J. Barnes, Samantha J. Ziegler, Ashutosh Mittal, Kelley W. Moremen, William S. York, Yannick J. Bomble, Maria J. Peña, Breeanna R. Urbanowicz
Xylans are a diverse family of hemicellulosic polysaccharides found in abundance within the cell walls of nearly all flowering plants. Unfortunately, naturally occurring xylans are highly heterogeneous, limiting studies of their synthesis and structure–function relationships. Here, we demonstrate that xylan synthase 1 from the charophyte alga Klebsormidium flaccidum is a powerful biocatalytic tool for the bottom-up synthesis of pure β-1,4 xylan polymers that self-assemble into microparticles in vitro. Using uridine diphosphate-xylose (UDP-xylose) and defined saccharide primers as substrates, we demonstrate that the shape, composition, and properties of the self-assembling xylan microparticles could be readily controlled via the fine structure of the xylan oligosaccharide primer used to initiate polymer elongation. Furthermore, we highlight two approaches for bottom-up and surface functionalization of xylan microparticles with chemical probes and explore the susceptibility of xylan microparticles to enzymatic hydrolysis. Together, these results provide a useful platform for structural and functional studies of xylans to investigate cell wall biosynthesis and polymer–polymer interactions and suggest possible routes to new biobased materials with favorable properties for biomedical and renewable applications.