Three highly acidic Equisetum XTHs differ from hetero-trans-β-glucanase in donor substrate specificity and are predominantly xyloglucan homo-transglucosylases.

Cell elongation Equisetum Hetero-trans-β-glucanase Heterologous expression Pichia pastoris Plant cell wall Xyloglucan endotransglucosylase

Journal

Journal of plant physiology
ISSN: 1618-1328
Titre abrégé: J Plant Physiol
Pays: Germany
ID NLM: 9882059

Informations de publication

Date de publication:
Aug 2020
Historique:
received: 13 08 2019
revised: 20 05 2020
accepted: 24 05 2020
pubmed: 17 6 2020
medline: 15 12 2020
entrez: 17 6 2020
Statut: ppublish

Résumé

Transglycanases are enzymes that remodel the primary cell wall in plants, potentially loosening and/or strengthening it. Xyloglucan endotransglucosylase (XET; EC 2.4.1.207), ubiquitous in land plants, is a homo-transglucanase activity (donor, xyloglucan; acceptor, xyloglucan) exhibited by XTH (xyloglucan endotransglucosylase/hydrolase) proteins. By contrast, hetero-trans-β-glucanase (HTG) is the only known enzyme that is preferentially a hetero-transglucanase. Its two main hetero-transglucanase activities are MLG : xyloglucan endotransglucosylase (MXE) and cellulose : xyloglucan endotransglucosylase (CXE). HTG is highly acidic and found only in the evolutionarily isolated genus of fern-allies, Equisetum. We now report genes for three new highly acidic HTG-related XTHs in E. fluviatile (EfXTH-A, EfXTH-H and EfXTH-I). We expressed them heterologously in Pichia and tested the encoded proteins' enzymic activities to determine whether their acidity and/or their Equisetum-specific sequences might confer high hetero-transglucanase activity. Untransformed Pichia was found to secrete MLG-degrading enzyme(s), which had to be removed for reliable MXE assays. All three acidic EfXTHs exhibited very predominantly XET activity, although low but measurable hetero-transglucanase activities (MXE and CXE) were also detected in EfXTH-H and EfXTH-I. We conclude that the extremely high hetero-transglucanase activities of Equisetum HTG are not emulated by similarly acidic Equisetum XTHs that share up to 55.5% sequence identity with HTG.

Identifiants

pubmed: 32544741
pii: S0176-1617(20)30100-0
doi: 10.1016/j.jplph.2020.153210
pii:
doi:

Substances chimiques

Plant Proteins 0
Glycosyltransferases EC 2.4.-
xyloglucan - xyloglucosyltransferase EC 2.4.1.207

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

153210

Informations de copyright

Copyright © 2020 Elsevier GmbH. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Competing Interest A patent application (WO2015044209) has been filed by BASF Agricultural Solutions Belgium NV and The University of Edinburgh for the use of hetero-transglycosylase. F.M., A.H., S.C.F., T.S. and C.H. are inventors.

Auteurs

Claire Holland (C)

The Edinburgh Cell Wall Group, Institute of Molecular Plant Sciences, The University of Edinburgh, Daniel Rutherford Building, The King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, UK.

Thomas J Simmons (TJ)

The Edinburgh Cell Wall Group, Institute of Molecular Plant Sciences, The University of Edinburgh, Daniel Rutherford Building, The King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, UK.

Frank Meulewaeter (F)

BASF Innovation Center Gent- Trait Research, Technologiepark-Zwijnaarde, 9052 Gent, Belgium.

Andrew Hudson (A)

Institute of Molecular Plant Sciences, The University of Edinburgh, Daniel Rutherford Building, The King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, UK.

Stephen C Fry (SC)

The Edinburgh Cell Wall Group, Institute of Molecular Plant Sciences, The University of Edinburgh, Daniel Rutherford Building, The King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, UK. Electronic address: s.fry@ed.ac.uk.

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Classifications MeSH