The History of Enzyme Evolution Embedded in Metabolism

Whereas phylogenetic reconstructions are a primary record of protein evolution, it is unknown whether the deep history of enzymes are encoded at higher levels of biological organization. Here, we demonstrate that the emergence and reuse history of enzymatic folds is embedded within the web of metabolite-cofactor-enzyme interdependencies that comprise biosphere-scale metabolic reaction networks. Using a simple network analysis approach, we reconstruct the relative ordering of enzymatic fold emergence and, where possible, the first reaction(s) that each enzymatic fold catalyzed. We find that a large majority of enzymatic folds were sufficient as independent additions to open new avenues for metabolic growth. The resulting network-based histories are broadly concordant with enzyme phyletic distribution in prokaryotes, a proxy for enzyme age. Our results suggest that the earliest enzyme-mediated metabolisms were enriched for /{beta} proteins, likely due to their strong association with cofactor utilization, and that -proteins preferentially emerge at later stages. The cradle-loop barrel, a member of the small {beta}-barrel metafold, is predicted to be the founding {beta}-fold, in agreement with analyses of ribosome structure. An examination of how the protein universe responded to the biological production of molecular oxygen reveals that the adaptation of existing enzymatic folds, not novel fold emergence, was the primary driver of metabolic evolution. This work presents a self-consistent model of metabolic and enzyme evolution, key progress towards integrating diverse perspectives into a unified history of protein evolution. Significance StatementEnzyme emergence is an ongoing process that began [~]4 billion years ago. Here, we show that the modern biosphere-scale network of metabolic reactions and enzymes is an archive of enzyme history independent from, but concordant with, phylogenetics. Based on this record, we predict the order of enzyme emergence from before the last universal common ancestor up until the biological production and metabolic utilization of molecular oxygen. We find that while /{beta} proteins dominated primitive enzyme-mediated metabolism, other folds -- including the cradle-loop barrel, which is a member of the small {beta}-barrel metafold -- were likely important early contributors. This study represents key progress towards building an internally consistent, joint history of metabolic reactions and enzymes.