G‑EnzyBeads for Biocatalysis Screening Facilitated by Volumetric Dosing

Screening catalysts is an essential step to identify more sustainable and efficient routes for the desired chemical transformations. While several concepts for streamlining high-throughput screenings are already present in chemocatalysis, biocatalysis requires the more challenging dosing of very small amounts of hygroscopic and electrostatic enzyme powders in large numbers to cover the catalyst space of interest. Within this work, the EnzyBeads technology, where enzymes are transiently bound to an inert, free-flowing support to allow dosing of tiny quantities of enzymes, was further developed and coupled with a parallel volumetric dosing approach in a 96-well format. A simplified coating procedure to coat glass beads, an inert carrier that was previously not usable due to inefficient adsorption, with the enzymes of interest was developed, which circumvents the use of resonance acoustic mixing and of the electrostatic polystyrene beads currently required in the status quo procedure. Using a custom-built 3D printable solid dispenser, named the 12X-Dosing Rack, the obtained enzyme-coated glass beads (G-EnzyBeads) can be dispensed in microtiter plates in a quick and reliable fashion to greatly facilitate biocatalysis screenings.