Fig. 2: Deep mutational scanning of DAOx expression and catalytic activity by EP-Seq.
From: Understanding activity-stability tradeoffs in biocatalysts by enzyme proximity sequencing
A Sorting gates for analysis of display levels. B Linear regression (Pearson r, two-tailed) between expression scores calculated from two biological replicates. C Sorting gates for catalytic activity screening. D Linear regression (Pearson r, two-tailed) between activity scores calculated from two biological replicates. E Linear regression (Pearson r, two-tailed) between variant surface display level measured in monogenic yeast culture vs. DMS expression fitness analyzed by EP-Seq for 12 DAOx single mutant variants. F Distribution of expression fitness effects measured by EP-Seq. Dashed lines represent the range of fitness score for synonymous variants. G Linear regression (Pearson r, two-tailed) between variant activity level measured in monogenic yeast culture via peroxidase assay (Amplex Red) vs. DMS activity fitness analyzed by EP-Seq for 12 DAOx single mutant variants. H Distribution of activity fitness effects measured by EP-Seq. Dashed lines represent the range of fitness score for synonymous variants. I Expression fitness scores for each variant represented as a heatmap. J Number of variants analyzed per position in the expression dataset, and secondary structure classification per position (PDB: 1C0P). K Activity fitness scores for each variant obtained by EP-Seq represented as a heatmap. L Number of variants analyzed per position in the activity dataset and secondary structure classification per position (PDB: 1C0P). Links to interactive and color blind accessible heatmaps can be found in the data availability statement section of the manuscript.
