Fig. 1: Schematic depicting the EP-Seq workflow.
From: Understanding activity-stability tradeoffs in biocatalysts by enzyme proximity sequencing
(Top) A pooled library of enzyme variants is displayed on yeast. (Left) The cell population is sorted into bins based on the expression level of the displayed enzyme. (Right) The pooled variant library is assayed for DAOx activity using a cascade peroxidase-mediated proximity labeling reaction with single cell fidelity and sorted into bins. The genetic composition of cells in the sorted bins is quantified via high-throughput sequencing and the distribution of each variant along the expression and activity axes is converted into a fitness score. Joint analysis of the two independent datasets provides insights into the effects of mutations on folding stability and activity of the enzyme.
