Hetero-trans-β-glucanase produces cellulose–1 xyloglucan covalent bonds in the cell walls of structural plant tissues and is stimulated by expansin

Klaus Herburger, Lenka Frankova, Martina Picmanova, Jai Wooi Loh, Marcos Valenzuela Ortega, Frank Meulewaeter, Andrew Hudson, Christopher French, Stephen Fry

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Current cell-wall models assume no covalent bonding between cellulose and hemicelluloses such as xyloglucan or mixed-linkage β-d-glucan (MLG). However, Equisetum hetero-trans-β-glucanase (HTG) grafts cellulose onto xyloglucan oligosaccharides (XGOs) – and, we now show, xyloglucan polysaccharide – in vitro, thus exhibiting CXE (cellulose:xyloglucan endotransglucosylase) activity. In addition, HTG also catalyzes MLG-to-XGO bonding (MXE activity). In this study, we explored the CXE action of HTG in native plant cell walls and tested whether expansin exposes cellulose to HTG by disrupting hydrogen bonds. To quantify and visualize CXE and MXE action, we assayed the sequential release of HTG products from cell walls pre-labeled with substrate mimics. We demonstrated covalent cellulose–xyloglucan bonding in plant cell walls and showed that CXE and MXE action was up to 15% and 60% of total transglucanase action, respectively, and peaked in aging, strengthening tissues: CXE in xylem and cells bordering intercellular canals and MXE in sclerenchyma. Recombinant bacterial expansin (EXLX1) strongly augmented CXE activity in vitro. CXE and MXE action in living Equisetum structural tissues potentially strengthens stems, while expansin might augment the HTG-catalyzed CXE reaction, thereby allowing efficient CXE action in muro. Our methods will enable surveys for comparable reactions throughout the plant kingdom. Furthermore, engineering similar hetero-polymer formation into angiosperm crop plants may improve certain agronomic traits such as lodging tolerance.
Original languageEnglish
Pages (from-to)1047-1062
Number of pages16
JournalMolecular Plant
Issue number7
Early online date3 May 2020
Publication statusPublished - 6 Jul 2020

Keywords / Materials (for Non-textual outputs)

  • cell wall
  • cellulose
  • Equisetum
  • hermicelluloses
  • hetero-trans-β-glucanase
  • hetero transglycosylation
  • in-situ localisation
  • plant morphology
  • xyloglucan


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