FIG 5.

The inhibitory effects of feruloyl amide are conserved in glucose fermentations. All experiments were performed similarly to xylose fermentations. (A) Anaerobic E. coli culture were grown on 1% glucose and treated with 5.5 mM feruloyl amide at mid-exponential phase (OD₆₀₀ of ∼0.4). Intracellular metabolites were collected at 0, 10, 30, 60, 120, 210, and 300 min after exposure to feruloyl amide and analyzed by LC-MS. The data represent the average of two biological replicates. (B) Anaerobic growth inhibition by 5.5 mM feruloyl amide. (C) Rapid PRPP accumulation after the feruloyl amide treatment. Similar to the observation in cells grown in xylose, PRPP rapidly increased upon exposure to 5.5 mM feruloyl amide. The accumulated PRPP was ¹³C labeled, indicating a metabolic bottleneck downstream of PRPP synthesis. Bars represent means of four biological replicates ± SEM. The fold change shown is nontreated control cells. (D) Isotopic tracers experiments using [¹³C]glucose also showed inhibition of ¹³C incorporation into purine and pyrimidine nucleotides.