Brushes of self-assembled nanotubes for temperature-responsive biocatalysis

Inspired by the elegant solutions that Nature has provided to control and promote specific site-reactions, my work presents an attempt to mimic filamentous biocatalytic interfaces. Brushes of self-assembled nanotubes with an enzymatic component (beta-lactamase) were prepared taking advantage of preexisting nanofabrication techniques, such as layer-by-layer and hard-templating. First, the effects of geometrical confinement and its consequences were investigated by comparison of (chitosan/beta-lactamase) multilayer film assembly on flat surfaces and in nanoporous membranes. In a second stage, polyelectrolyte nanotubes with controlled dimensions were prepared in nanoporous membranes and further anchored on a surface by chemical crosslinking to obtain brushes of nanotubes. The kinetic studies revealed the presence of active enzyme in the brushes and enhanced activity preservation when beta-lactamase was deposited as the inner layers of the nanotubes. As a final step, a variety of thermo-responsive coatings with different architectures were tested to control substrate diffusion on top of beta-lactamase-based multilayer films. The integration of stable thermo-responsive elements was proven, although further experiments are required to control biocatalysis with additional layers and using more complex mechanisms, such as coupled thermal and mechanical responses. Knowing that there are more challenges to face before reaching optimum nanotube brushes and apply them for controlled biocatalysis, this study contributes with some elements that may pave the way towards the integration of different techniques for the fabrication of complex biocatalytic nanostructures.