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The main goal of the project is to develop a tool-box for the fabrication of hierarchically organized heterogeneous multi-functional biocatalysts.

As proof of the concept, we have designed two artificial cascades for the synthesis of omega-aminoacids (omega-AA) using diols and bio-oils as renewable raw materials; both cascades will be based on 4 different functional modules (A-D). We will assemble the different enzymes forming modules A-D into protein scaffolds based on the consensus tetratricopeptide repeats (CTPR) to endow such functional modules with nanometric proximity. We will construct a set of plasmid encoding chimeric proteins with functional (enzyme) and structural (scaffold brick) domains in order to assemble 4 functional modules that can be readily immobilized on solid carriers to create a battery of modular heterogeneous biocatalysts. The immobilized functional modules can be combined ad hoc to catalyze one or the other synthetic scheme depending on the available raw materials. We will demonstrate the applicability of the immobilized functional modules by demonstrating and scaling-up the synthesis of long and short -AA. Finally the fabrication of one set of plasmids and one kit for solid-phase and tailor-made assembly of different enzyme-scaffold units will be demonstrated and validated at industrial level.