title: An enzyme activity capable of endotransglycosylation of heteroxylan polysaccharides is present in plant primary cell walls.
abstract: Heteroxylans in the plant cell wall have been proposed to have a role analogous to that of xyloglucans or heteromannans, forming growth-restraining networks by interlocking cellulose microfibrils. A xylan endotransglycosylase has been identified that can transglycosylate heteroxylan polysaccharides in the presence of xylan-derived oligosaccharides. High activity was detected in ripe fruit of papaya (Carica papaya), but activity was also found in a range of other fruits, imbibed seeds and rapidly growing seedlings of cereals. Xylan endotransglycosylase from ripe papaya fruit used a range of heteroxylans, such as wheat arabinoxylan, birchwood glucuronoxylan and various heteroxylans from dicotyledonous primary cell walls purified from tomato and papaya fruit, as donor molecules. As acceptor molecules, the enzyme preferentially used xylopentaitol over xylohexaitol or shorter-length acceptors. Xylan endotransglycosylase was active over a broad pH range and could perform transglycosylation reactions up to 55 °C. Xylan endotransglycosylase activity was purified from ripe papaya fruit by ultrafiltration and cation exchange chromatography. Highest endotransglycosylase activity was identified in fractions that also contained high xylan hydrolase activity and correlated with the presence of the endoxylanase CpaEXY1. Recombinant CpaEXY1 protein transiently over-expressed in Nicotiana benthamiana leaves showed both endoxylanase and xylan endotransglycosylase activities in vitro, suggesting that CpaEXY1 is a single enzyme with dual activity in planta. Purified native CpaEXY1 showed two- to fourfold higher endoxylanase than endotransglycosylase activity, suggesting that CpaEXY1 may act primarily as a hydrolase. We propose that xylan endotransglycosylase activity (like xyloglucan and mannan endotransglycosylase activities) could be involved in remodelling or re-arrangement of heteroxylans of the cellulose-non-cellulosic cell wall framework.
authors.affiliation: The New Zealand Institute for Plant and Food Research Limited, Mount Albert Research Centre, Private Bag 92169, Auckland 1142, New Zealand.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Johnston
authors.forename: Roneel
authors.initials: R
authors.lastname: Prakash
authors.forename: Nancy J
authors.initials: NJ
authors.lastname: Chen
authors.forename: Monto H
authors.initials: MH
authors.lastname: Kumagai
authors.forename: Helen M
authors.initials: HM
authors.lastname: Turano
authors.forename: Janine M
authors.initials: JM
authors.lastname: Cooney
authors.forename: Ross G
authors.initials: RG
authors.lastname: Atkinson
authors.forename: Robert E
authors.initials: RE
authors.lastname: Paull
authors.forename: Roshan
authors.initials: R
authors.lastname: Cheetamun
authors.forename: Antony
authors.initials: A
authors.lastname: Bacic
authors.forename: David A
authors.initials: DA
authors.lastname: Brummell
authors.forename: Roswitha
authors.initials: R
authors.lastname: Schröder
mesh_terms: D000595:Amino Acid Sequence; D029441:Carica / Q000201:enzymology / Q000378:metabolism; D002473:Cell Wall / Q000201:enzymology* / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases / Q000235:genetics / Q000378:metabolism; D005638:Fruit / Q000201:enzymology / Q000378:metabolism; D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000378:metabolism; D007700:Kinetics; D018551:Solanum lycopersicum / Q000201:enzymology / Q000378:metabolism; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000235:genetics; D010940:Plant Proteins / Q000378:metabolism*; D010944:Plants / Q000201:enzymology* / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature; D014026:Nicotiana / Q000235:genetics; D014990:Xylans / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D014990:Xylans; D016695:Glycosyltransferases; D006867:Hydrolases; D043364:Endo-1,4-beta Xylanases