Biocatalytic programming of protocell-embodied logic gates and circuits

The construction of biomimetic agents capable of generating precise outcomes in response to specific molecular inputs is a central challenge for the development of programmable synthetic cells with integrated biomimetic functions. Here, we harness acoustic standing waves to generate periodic microarrays of enzyme-encoded coacervate microdroplets for the implementation of embodied enzyme logic circuits (EELCs). We describe a range of biocatalytic communication channels capable of performing localized and distributed Boolean logic functions in single or segregated populations of model protocells by using a range of molecular inputs, fluorescence or hydrogelation outputs, and programmable response dynamics. To implement long-range collective signal processing, we integrate EELC modules across spatially segregated protocell populations to generate distributed time-regulated logic operations involving negative feedback, pulse generation, and redirected output-input connectivity. Our results provide a step toward the non-DNA programming of model protocell communication and computational networks for miniaturized autonomous sensing devices capable of chemical-based information processing.