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Planta

, Volume 237, Issue 1, pp 173–187 | Cite as

An enzyme activity capable of endotransglycosylation of heteroxylan polysaccharides is present in plant primary cell walls

  • Sarah L. Johnston
  • Roneel Prakash
  • Nancy J. Chen
  • Monto H. Kumagai
  • Helen M. Turano
  • Janine M. Cooney
  • Ross G. Atkinson
  • Robert E. Paull
  • Roshan Cheetamun
  • Antony Bacic
  • David A. Brummell
  • Roswitha SchröderEmail author
Original Article

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.

Keywords

Cell wall Endoxylanase Heteroxylan Papaya Xylan endotransglycosylase 

Abbreviations

CTAB

Cetyltrimethylammonium bromide

WAX

Wheat arabinoxylan

PAHBAH

p-Hydroxybenzoic acid hydrazide

XET

Xyloglucan endotransglycosylase

XTH

Xyloglucan endotransglycosylase/hydrolase

Xyl2-ol

Xylobiitol

Xyl3-ol

Xylotriitol

Xyl4-ol

Xylotetraitol

Xyl5-ol

Xylopentaitol

Xyl6-ol

Xylohexaitol

Notes

Acknowledgments

We thank Erin O’Donoghue, William Laing and Robert Schaffer for comments on the manuscript. We are indebted to Lionel M. Hill (John Innes Centre, Norwich, England) for his advice with the enzyme kinetics. We also thank Cherie Walsh, ARC Centre of Excellence in Plant Cell Walls, The University of Melbourne, for technical assistance with cell wall methylation analyses. Antony Bacic acknowledges the support of the ARC Centre of Excellence in Plant Cell Walls. This project was funded by the Ministry of Business Innovation and Employment, New Zealand (contract C11X1007).

Supplementary material

425_2012_1766_MOESM1_ESM.pdf (94 kb)
Table S1 Peptide fragments in fraction 16 eluted from the MonoS column (PDF 94.1 kb)
425_2012_1766_MOESM2_ESM.pdf (264 kb)
Table S2 Predicted xylanase proteins and open reading frames in the papaya genome (PDF 263 kb)
425_2012_1766_MOESM3_ESM.pdf (172 kb)
Fig. S1 Analysis of the recombinant protein products produced in Nicotiana benthamiana leaves using viral vector TTOSA1-XYL (PDF 172 kb)
425_2012_1766_MOESM4_ESM.pdf (257 kb)
Fig. S2 2D-gel electrophoresis and gel blot of papaya fruit proteins (PDF 256 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Sarah L. Johnston
    • 1
  • Roneel Prakash
    • 1
  • Nancy J. Chen
    • 2
  • Monto H. Kumagai
    • 2
    • 6
  • Helen M. Turano
    • 2
  • Janine M. Cooney
    • 3
  • Ross G. Atkinson
    • 1
  • Robert E. Paull
    • 2
  • Roshan Cheetamun
    • 4
  • Antony Bacic
    • 4
  • David A. Brummell
    • 5
  • Roswitha Schröder
    • 1
    Email author
  1. 1.The New Zealand Institute for Plant and Food Research LimitedMount Albert Research CentreAucklandNew Zealand
  2. 2.Department of Tropical Plant and Soil SciencesUniversity of Hawaii at ManoaHonoluluUSA
  3. 3.The New Zealand Institute for Plant and Food Research LimitedRuakura Research CentreHamiltonNew Zealand
  4. 4.ARC Centre of Excellence in Plant Cell Walls, School of BotanyUniversity of MelbourneMelbourneAustralia
  5. 5.The New Zealand Institute for Plant and Food Research LimitedFood Industry Science CentrePalmerston NorthNew Zealand
  6. 6.XtremeSignPost Inc.LeucadiaUSA

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