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https://hdl.handle.net/2440/60683
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Type: | Journal article |
Title: | Polysaccharide microarrays for high-throughput screening of transglycosylase activities in plant extracts |
Author: | Kosik, O. Auburn, R. Russell, S. Stratilova, E. Garajova, S. Hrmova, M. Farkas, V. |
Citation: | Glycoconjugate Journal, 2010; 27(1):79-87 |
Publisher: | Kluwer Academic Publ |
Issue Date: | 2010 |
ISSN: | 0282-0080 1573-4986 |
Statement of Responsibility: | Ondřej Kosík, Richard P. Auburn, Steven Russell, Eva Stratilová, Soňa Garajová, Maria Hrmova and Vladimír Farkaš |
Abstract: | Polysaccharide transglycosylases catalyze disproportionation of polysaccharide molecules by cleaving glycosidic linkages in polysaccharide chains and transferring their cleaved portions to hydroxyl groups at the non-reducing ends of other polysaccharide or oligosaccharide molecules. In plant cell walls, transglycosylases have a potential to catalyze both cross-linking of polysaccharide molecules and grafting of newly arriving polysaccharide molecules into the cell wall structure during cell growth. Here we describe a polysaccharide microarray in form of a glycochip permitting simultaneous high-throughput monitoring of multiple transglycosylase activities in plant extracts. The glycochip, containing donor polysaccharides printed onto nitrocellulose-coated glass slides, was incubated with crude plant extracts, along with a series of fluorophore-labelled acceptor oligosaccharides. After removing unused labelled oligosaccharides by washing, fluorescence retained on the glycochip as a result of transglycosylase reaction was detected with a standard microarray scanner. The glycochip assay was used to detect transglycosylase activities in crude extracts from nasturtium (Tropaeolum majus) and mouse-ear cress (Arabidopsis thaliana). A number of previously unknown saccharide donor-acceptor pairs active in transglycosylation reactions that lead to the formation of homo- and hetero-glycosidic conjugates, were detected. Our data provide experimental support for the existence of diverse transglycosylase activities in crude plant extracts. |
Keywords: | Glycochip Microarray Oligosaccharides Plant cell wall Transglycosylation XET |
Rights: | Copyright © 2010, Springer Science+Business Media, LLC |
DOI: | 10.1007/s10719-009-9271-8 |
Published version: | http://dx.doi.org/10.1007/s10719-009-9271-8 |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest |
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