Cell-free biosynthesis allows for synthesizing pharmacologically as well as technically relevant proteins and enzymes in an energy-economically optimized environment – especially those enzymes and proteins that are cytotoxic and thus hardly accessible by conventional, cell-based methods. This talk points out how microscopic polymer materials contribute to the development of synthetic platforms for studying, manipulating and optimizing cell-inspired syntheses and functions.
To manufacture polymer materials that serve as robust synthesis and analysis microenvironments with defined parameter space, innovative processing technologies are required to control physicochemical polymer material properties and mechanics on the micrometer-scale. On this account, we utilize two methods: droplet microfluidics to fabricate polymer microgels swollen in water, and high-resolution additive manufacturing based on micro-stereolithography (µSL) to process polymers with micron-scale precision in a solvent-based (hydrogels) or solvent-free (mass polymerization) fashion.
By combining these two methods, experimental platforms emerge for understanding and optimizing cellular functions (e.g., protein synthesis and enzymatic cascade reactions) in an artificial environment that still reflects key aspects of cellular life (e.g., diffusivity and spatiotemporal compartmentalization). Exemplarily, we immobilize sensitive enzymes for polyketide synthesis in microgels, whose polymer matrix is optimized regarding porosity and hydrophobicity to preserve enzyme conformation and activity. By loading these microgels into 3D-printed flow cells, we manipulate experimental conditions with spatiotemporal control within microseconds and micrometers, respectively.