Biocatalytic process engineering with the use of 3D printing technology

Abstract

This diploma thesis investigates the integration of 3D printing technology with biocatalysis to develop advanced microfluidic systems for enzyme immobilization and activity assessment. Research and experimental work were conducted at the Institute of Technical Chemistry, Leibniz University Hannover, Germany, in the Biocatalysis and Bioprocessing research group. This study explores the design and construction of microfluidic reactors using cutting-edge 3D printing techniques in order to optimize enzymatic processes. Our goal is to improve the stability and effectiveness of enzyme immobilization by incorporating surface chemistry techniques like glutaraldehyde cross-linking and coatings with polydopamine and polyethyleneimine. The primary objectives include the development of microfluidic systems for selective enzyme immobilization, assessment of biocatalytic performance under continuous flow conditions, and identification of optimal materials and 3D printing methodologies. Utilizing Fused Deposition Modeling and Stereolithography printing technologies in conjunction with enzyme expression and immobilization techniques, precise microfluidic devices are created for effective enzymatic reactions. These prototypes serve as effective screening platforms and can be parallelized to enhance production capacity, which will help with sustainable chemistry and biotechnology applications. As an example of how different 3D printing materials can be used for biotechnological applications like enzyme immobilization, this work emphasizes on how important microfluidic chip-based assays are for effective and quick screening because of their low reagent amounts, affordability, and rapid throughput. In particular, unspecific peroxygenase from Agrocybe aegerita (AaeUPO) was successfully immobilized on surface functionalized 3D-printed microfluidic chips made of different materials, as confirmed by a model activity assay. This was the first time that 3D-printed microchips were used for AaeUPO immobilization, and the enzyme demonstrated high immobilization and activity yields across various materials and geometries, highlighting the potential for development of a universal immobilization approach.