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    Building an Artificial Plant Cell Wall on a Lipid Bilayer by Assembling Polysaccharides and Engineered Proteins

    Year: 2022

    Journal: ACS Synth. Biol., Volume 11, OCT 21, page 3516–3528

    Authors: Notova, Simona; Cannac, Nathan; Rabagliati, Luca; Touzard, Maeva; Mante, Josselin; Navon, Yotam; Coche-Guerente, Liliane; Lerouxel, Olivier; Heux, Laurent; Imberty, Anne

    Organizations: European Union Horizon 2020 Research and Innovation Program under the Marie Sklodowska-Curie grant agreement synBIOcarb [814029]; Glyco@Alps program [ANR-15-IDEX-0002, ANR-17-EURE-0003]

    Keywords: synthetic biology; protein engineering; lectin; carbohydrate-binding protein; plant cell wall

    The cell wall constitutes a fundamental structural component of plant cells, providing them with mechanical resistance and flexibility. Mimicking this wall is a critical step in the conception of an experimental model of the plant cell. The assembly of cellulose/hemicellulose in the form of cellulose nanocrystals and xyloglucans as a representative model of the plant cell wall has already been mastered; however, these models lacked the pectin component. In this work, we used an engineered chimeric protein designed for bridging pectin to the cellulose/ hemicellulose network, therefore achieving the assembly of complete cell wall mimics. We first engineered a carbohydrate-binding module from Ruminococcus flavefaciens able to bind oligogalacturonan, resulting in high-affinity polygalacturonan receptors with Kd in the micromolar range. A Janus protein, with cell wall gluing property, was then designed by assembling this carbohydrate-binding module with a Ralstonia solanacearum lectin specific for fucosylated xyloglucans. The resulting supramolecular architecture is able to bind fucose-containing xyloglucans and homogalacturonan, ensuring high affinity for both. A two-dimensional assembly of an artificial plant cell wall was then built first on synthetic polymer and then on the supported lipid bilayer. Such an artificial cell wall can serve as a basis for the development of plant cell mechanical models and thus deepen the understanding of the principles underlying various aspects of plant cells and tissues.
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