title: Building custom polysaccharides in vitro with an efficient, broad-specificity xyloglucan glycosynthase and a fucosyltransferase.
abstract: The current drive for applications of biomass-derived compounds, for energy and advanced materials, has led to a resurgence of interest in the manipulation of plant polymers. The xyloglucans, a family of structurally complex plant polysaccharides, have attracted significant interest due to their intrinsic high affinity for cellulose, both in muro and in technical applications. Moreover, current cell wall models are limited by the lack of detailed structure-property relationships of xyloglucans, due to a lack of molecules with well-defined branching patterns. Here, we have developed a new, broad-specificity "xyloglucan glycosynthase", selected from active-site mutants of a bacterial endoxyloglucanase, which catalyzed the synthesis of high molar mass polysaccharides, with complex side-chain structures, from suitable glycosyl fluoride donor substrates. The product range was further extended by combination with an Arabidopsis thaliana α(1→2)-fucosyltransferase to achieve the in vitro synthesis of fucosylated xyloglucans typical of dicot primary cell walls. These enzymes thus comprise a toolkit for the controlled enzymatic synthesis of xyloglucans that are otherwise impossible to obtain from native sources. Moreover, this study demonstrates the validity of a chemo-enzymatic approach to polysaccharide synthesis, in which the simplicity and economy of glycosynthase technology is harnessed together with the exquisite specificity of glycosyltransferases to control molecular complexity.
authors: Division of Glycoscience, School of Biotechnology, Royal Institute of Technology (KTH), 106 91 Stockholm, Sweden.
authors: Oliver
authors: O
authors: Spadiut
authors: Farid M
authors: FM
authors: Ibatullin
authors: Jonelle
authors: J
authors: Peart
authors: Fredrika
authors: F
authors: Gullfot
authors: Carlos
authors: C
authors: Martinez-Fleites
authors: Marcus
authors: M
authors: Ruda
authors: Chunlin
authors: C
authors: Xu
authors: Gustav
authors: G
authors: Sundqvist
authors: Gideon J
authors: GJ
authors: Davies
authors: Harry
authors: H
authors: Brumer
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