High-Throughput Screening of Drug-Resistant Viral Protease and Acid-Tolerant Glutenase using Droplet Microfluidics
Enzymes are functionalized proteins that act as catalysts, accelerating chemical reactions in biological systems. The enzyme evolution, which refers to the enzymes change in their sequence, brings the enzyme benefits such as tolerances to high temperature, acid, base and strong salinity, but sometimes the declines on targeting, binding and catalyzing efficiency. Our goal is to harness the potential of high-performing enzyme mutants while also elucidating the underlying mechanisms and strategies to improve less efficient enzymes. Here, we develop a high-throughput screening assay that combines droplet microfluidics with direct enzyme evolution. We produce a mutation library over 106 and screen out the enzyme mutants we are interested in. Specifically, we have applied our technique on the screening of drug resistant mutants of the SARS-CoV-2 (virus caused the COVID19) Main-protease (Junming Lao, Karla Durdic). Besides, we have promoted the evolution and identification of gastric acid-tolerant glutenases, offering a potential therapeutic avenue for individuals with gluten allergies based on this technology (Xinge Zhang, Junming Lao).
