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Biochemical characterization of family 43 glycosyltransferases in the Populus xylem: challenges and prospects

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Winzell, Anders, Guerriero, Gea, Aspeborg, Henrik, Wang, Yiqiang, Rajangam, Alex S., Teeri, Tuula T. and Ezcurra, Ines (2010) Biochemical characterization of family 43 glycosyltransferases in the Populus xylem: challenges and prospects. In: Colloquium on Green Chemistry in Plants and Microorganisms, Japanese Soc Promot Sci, Stockholm, Sweden, May 25, 2009. Published in: Plant Biotechnology, Vol.27 (No.3). pp. 283-288.

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Official URL: http://www.jstage.jst.go.jp/article/plantbiotechno...

Abstract

Wood formation is a biological process of great economical importance. Genes active during the secondary cell wall formation of wood fibers from Populus tremulaxtremuloides were previously identified by expression profiling through microarray analyses. A number of these genes encode glycosyltransferases (GTs) with unknown substrate specificities. Here we report heterologous expression of one of these enzymes, PttGT43A, a putative IRREGULAR XYLEM9 (IRX9) homologue. Expression trials in Pichia pastoris and insect cells revealed very low levels of accumulation of immunoreactive PttGT43A, whereas transient expression in Nicotiana benthamiana leaves by Agrobacterium infiltration (agroinfiltration) using a viral vector produced substantial amounts of protein that mostly precipitated in the crude pellet. Agroinfiltration induced weak endogenous xylosyltransferase activity in microsomal extracts, and transient PttGT43A expression further increased this activity, albeit only to low levels. PttGT43A may be inactive as an individual subunit, requiring complex formation with unknown partners to display enzymatic activity. Our results suggest that transient co-expression in leaves of candidate subunit GTs may provide a viable approach for formation of an active xylan xylosyltransferase enzymatic complex.

Item Type: Conference Item (UNSPECIFIED)
Subjects: T Technology > TP Chemical technology
S Agriculture > SB Plant culture
Divisions: Faculty of Science > Life Sciences (2010- ) > Warwick HRI (2004-2010)
Journal or Publication Title: Plant Biotechnology
Publisher: Japanese Society for Plant Cell and Molecular Biology
ISSN: 1342-4580
Date: July 2010
Volume: Vol.27
Number: No.3
Number of Pages: 6
Page Range: pp. 283-288
Status: Not Peer Reviewed
Publication Status: Published
Title of Event: Colloquium on Green Chemistry in Plants and Microorganisms
Type of Event: Conference
Location of Event: Japanese Soc Promot Sci, Stockholm, Sweden
Date(s) of Event: May 25, 2009
URI: http://wrap.warwick.ac.uk/id/eprint/5492

Data sourced from Thomson Reuters' Web of Knowledge

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