Dissecting enzyme function with microfluidic-based deep mutational scanning

TLDR

Enzymes, powerful biological catalysts, enable industrial chemical production and disease treatment, yet our limited understanding of catalysis hampers the design of new enzymes. The study aims to develop a method that uses unbiased empirical analysis to dissect enzyme function, facilitating the design of enzymes with tailored chemical functions. The method employs unbiased empirical analysis to map how amino‑acid changes affect enzyme activity. Comprehensive mapping of amino‑acid substitutions reveals the interactions shaping the enzyme function landscape and uncovers new biochemical mechanisms that enhance custom biocatalyst engineering.

Abstract

Significance As powerful biological catalysts, enzymes can solve challenging problems that range from the industrial production of chemicals to the treatment of human disease. The ability to design new enzymes with tailor-made chemical functions would have a far-reaching impact. However, this important capability has been limited by our cursory understanding of enzyme catalysis. Here, we report a method that uses unbiased empirical analysis to dissect the molecular basis of enzyme function. By comprehensively mapping how changes in an enzyme’s amino acid sequence affect its activity, we obtain a detailed view of the interactions that shape the enzyme function landscape. Large, unbiased analyses of enzyme function allow the discovery of new biochemical mechanisms that will improve our ability to engineer custom biocatalysts.