Fig. 6: TESEC supports screening for diverse metabolic targets.
From: Low-cost anti-mycobacterial drug discovery using engineered E. coli
a–d TESEC strains for the indicated metabolic targets grown on defined medium with the essential enzyme induced with a range of arabinose concentrations. The indicated low (L) and high (H) arabinose concentrations were selected for differential screening. Data are presented as the mean and CI95 of six biological replicates. e–h Differential screening of the Prestwick library was performed at both low and high arabinose concentrations in triplicate. Hits were selected as compounds with median differential OD measurements of 0.1 and SSMD scores >5. i, j Hit compounds identified for targets Asd, and DapB. No significant hits were identified for CysH or TrpD. Colored bars indicate mean growth under high induction, gray bars at low induction. Individual data points are shown for three biological replicates. k Molecular weight and hydrophobicity scores for identified hit compounds (Asd, blue; DapB, purple) and for all annotated antibacterials in the Prestwick library (beige). Hydrophobicity scores were estimated with the alvaMolecule software package. l–p Chemical-genetic validation profiles show TESEC grows as a function of both drug dose and TESEC target induction level. With the exception of diethylstilbestrol, hit compounds showed the characteristic diagonal pattern indicating that drug sensitivity depended on target expression level. q–u Individual dose–response curves extracted from the chemical-genetic profiles for a range of arabinose concentrations and the indicated drug dose. Except diethylstilbestrol, drug-treated TESEC strains showed improved growth at higher arabinose induction (green lines). Wild-type controls, in which target expression was not controlled by arabinose, did not show improved growth (gray lines). Data are presented as the mean and CI95 for three biological replicates. Source data are provided as a Source Data file.
