Urgently needed innovations in sustainability that are useful across geographies and economies and can operate at scale pose a technological opportunity. Toward these innovations, we are developing integrated molecular, systems, and process level capabilities with biological parts to define how we strategically use waste as substrates key to our energy future. Specifically, we are advancing the scope, scale, and impact of cell-free expression systems to do this. The foundational principle is that we can conduct precise, complex biomolecular transformations in purified protein or crude cell lysate systems. This approach circumvents mechanisms evolved to facilitate species survival, bypasses limitations on molecular transport across the cell wall, and provides a significant departure from traditional, cell-based processes. In this talk, I will describe our high-throughput, machine learning-guided approach to enable exploration of protein fitness landscapes across multiple regions of chemical space for forward design of protein activities. By evaluating tens of thousands of unique protein sequences, we have developed synthetic biology solutions to several sustainability challenges. I will highlight our development of a toolbox of biocatalysts for green production of high-value pharmaceuticals, a synthetic metabolic pathway to improve C1 utilization, and protein-based sensors for environmental contaminants. This work demonstrates that cell-free systems offer an agile, modular, and powerful engineering strategy that can enable new biotechnologies capable of impacting all industries, people, and the planet.
