Evolution-based design of chorismate mutase enzymes

The rational design of enzymes is an important goal for both fundamental and practical reasons. Here, we describe a design process in which we learn the constraints for specifying proteins purely from evolutionary sequence data, build libraries of synthetic genes, and test them for activity in vivo using a quantitative complementation assay. For chorismate mutase, a key enzyme in the biosynthesis of aromatic amino acids, we demonstrate the design of natural-like catalytic function with substantial sequence diversity. Further optimization focuses the generative model towards function in a specific genomic context. The data show that sequence-based statistical models suffice to specify proteins and provide access to an enormous space of synthetic functional sequences. This result provides a foundation for a general process for evolution-based design of artificial proteins. One-sentence summaryAn evolution-based, data-driven engineering process can build synthetic functional enzymes.