Fig. 7

Reverse engineering strategy for the 12 mutated genes. a Growth conditions of W3110 in the presence of 1 g/L salicylic acid (SA) over 24 h following the knockout of each of the 12 genes in W3110K-4. b Construction of plasmids for the 12 mutated genes using the medium-copy plasmid pACYCDuet, followed by their introduction into W3110K-4 to observe growth conditions and salicylic acid (SA) production yields. c Phage resistance profiles of W3110, W3110K-2, and W3110K-4. For the phage resistance assays, exponentially growing cultures (OD600 ≈ 0.6) were subjected to a tenfold dilution with PBS. A 2 μL aliquot of the diluted suspension was then pre-spotted onto LB agar plates. After allowing the surface to dry slightly, high-concentration phage suspensions were repeatedly spot-inoculated onto the E. coli area using sterile toothpicks to facilitate efficient phage–host interaction