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

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

From: Genome-wide mapping of mutations at single-nucleotide resolution for protein, metabolic and genome engineering

Supplementary Figure 13

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.

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