WP3: Enzyme immobilization

Objectives: Development of efficient ATA immobilization approach. Production of a hybrid catalyst of high catalytic efficiency.

Description of Work: Τhe ATAs selected in WP2 will be expressed, purified and covalently and/or non-covalently immobilized on the GO selected in WP1. The catalytic behavior (in terms of catalytic activity, stability and reusability) and the structural implications of the immobilization will be investigated. For comparison purposes, the non-optimized ATAs (prior to the protein engineering effort) will be immobilized on GO with the same processes, to identify if the advantageous profile of the free variants is maintained when immobilized, and if the immobilization has different effects on the ATAs. Acetophenone will be used as a benchmark substrate for the biocatalytic characterization (only of the enzymatic step of the cascade), as well as some compounds selected in WP1.