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    Supported Supramolecular Hydrogel Nanoarchitectonics for Tunable Biocatalytic Flow Activity

    Year: 2024

    Authors: More, Shahaji H.; Runser, Jean-Yves; Ontani, Aymeric; Fores, Jennifer Rodon; Carvalho, Alain; Blanck, Christian; Serra, Christophe A.; Schmutz, Marc; Schaaf, Pierre; Jierry, Loïc

    Keywords: biocatalytic flow process; enzyme-assisted self-assembly; peptide-based nanofiber; polyelectrolyte multilayer film; supramolecular hydrogel

    Enzymatically-active polyelectrolyte multilayers containing n layers of phosphatase (APn-PEM) induce the formation of supported biocatalytic supramolecular hydrogels when brought in contact with the precursor tripeptide Fmoc-FFpY (Fmoc = N-fluorenylmethyloxycarbonyl; F = Phenylalanine; Y = Tyrosine; p = phosphate group). APn-PEM triggers the spatially-localized hydrogelation reaching 2, 17 and 350 µm of thickness for n = 1, 2 and 3, respectively. As observed by cryo scanning electron microscopy, a dense nanofibrous network underpinning the hydrogel shows parallelly orientated Fmoc-FFY peptide-based fibrils, perpendicular to the substrate. For the gel generated by the AP3-PEM, fluorescence confocal microscopy images show that during the peptide self-assembly, some enzymes are distributed in the hydrogel, preferentially located in few dozens of micrometers above the substrate. In addition, a self-assembly growth rate of 5 µm min−1 is determined when the hydrogelation starts. Through transmission electron microscopy immuno-labelling experiments on self-assemblies generated in solution, we observe that AP are decorating the Fmoc-FFY nanofibers. It is observed both a long-term stability and a higher biocatalytic activity of the so AP-encapsulated hydrogel compared to the bare APn-PEM. This bioactivity can be tuned by the number n in batch and under continuous flow conditions. To illustrate the versatility of this enzyme-supported strategy, multi-catalytic transformations in continuous flow conditions have been successfully carried out using supported supramolecular hydrogel.
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