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The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression

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Abstract

The cell wall polymer lignin is believed to be condensed by specific cell wall-localized oxidoreductases. In many plants species, including poplar, the peroxidase-directed oxidation of the lignin analogue syringaldazine (SYR) has been localized to cells that undergo secondary wall formation, a process that includes lignification. As a first step to analyse the corresponding peroxidases, we have isolated previously two anionic isoenzymes (PXP 3-4 and PXP 5) from poplar xylem (Populus trichocarpa), which use SYR as a substrate. Here, we demonstrate that these enzymes are responsible for the visualized SYR oxidation in the developing xylem. The cDNA that corresponds to PXP 3-4 was isolated and the deduced protein was found closely related to the other SYR-oxidizing peroxidase PXP 5 (ca. 98% of identity). PXP 3-4 was expressed in a baculovirus expression system yielding high levels of active peroxidase (3 mg/l medium). The heterologously produced protein showed characteristics similar to those of the corresponding protein from poplar xylem (enzymatic properties, isoelectric point, and migration in a native gel). PXP 3-4 was expressed in the stem and in the root xylem. The data demonstrate that PXP 3-4 (and/or PXP 5) are present in differentiating xylem, supporting a function in secondary cell wall formation.

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Authors and Affiliations

  1. Vakgroep Moleculaire Genetica, Departement Plantengenetica, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB), Universiteit Gent, K.L. Ledeganckstraat 35, 9000 , Gent, Belgium

    Jørgen Holst Christensen, Antje Rohde, Wilson Ardiles Diaz, Guy Bauw, Marc Van Montagu & Wout Boerjan

  2. Plant Biochemistry and Physiology, University of Geneva, Place de l'Université 3, 1211, Genève 4, Switzerland

    Serge Overney & Patrice Simon

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  1. Jørgen Holst Christensen
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  2. Serge Overney
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  3. Antje Rohde
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  4. Wilson Ardiles Diaz
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  7. Marc Van Montagu
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Holst Christensen, J., Overney, S., Rohde, A. et al. The syringaldazine-oxidizing peroxidase PXP 3-4 from poplar xylem: cDNA isolation, characterization and expression. Plant Mol Biol 47, 581–593 (2001). https://doi.org/10.1023/A:1012271729285

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