Supplementary Figure 13: Validation of Crp S28P mutation for furfural or thermal tolerance.

a) Crystal structure of the Crp regulatory protein with variants identified by furfural selection highlighted in red (PDB ID 3N4M). A number of the CREATE designs targeting residues near the cyclic-AMP binding site (aa. 28-30, 65) of this regulator were highly enriched in minimal media selections for furfural or thermal tolerance, suggesting that these mutations may enhance E. coli growth in minimal media under a variety of stress conditions. b) Validation the Crp S28P mutant identified in 2 g/L furfural selections in M9 media. This mutant was reconstructed as described for AcrB T60S in Fig. S8.