Fig. 5: MA production via rational pathway engineering and DOL-mediated optimization.

a Design of a synthetic consortium that upcycles PET hydrolysate into MA. The consortium (TC-EM) consists of Pp-TC and Pp-EM derived from Pp-T and Pp-E, respectively. Involving two layers of DOL, the strains were divided to specialize in TPA (Pp-TC) and EG (Pp-EM) consumptions and to enable the TPA-to-CAT (Pp-TC) and CAT-to-MA (Pp-EM) conversions. b Temporal profiles of TPA fermentation by Pp-TC that yields CAT. c Temporal profiles of EG fermentation by Pp-EM that upgrades CAT into MA. d Design of a single strain (Pp-TEM) that produces MA from PET hydrolysate. e Temporal profiles of mixed TPA and EG fermentation by Pp-TEM that converts TPA and EG into MA. f Production optimization by modulating the initial strain inoculation ratio of the TC-EM consortium. g Temporal profiles of PET hydrolysate fermentation by the TC-EM consortium with a 1:5 initial ratio. h Comparison of the final MA production by the TC-EM consortium with different initial ratios and Pp-TEM. i Colors of the TC-EM co-culture and Pp-TEM monoculture through the course of hydrolysate fermentation. j Production optimization by altering the lag time between Pp-TC and Pp-EM inoculations. As illustrated, Pp-TC was inoculated after Pp-EM upon different durations of delay. k Comparison of final products and remnant substrates of the hydrolysate fermentation by the TC-EM consortium with varied inoculation lag times. For panels a and d, blue text indicates overexpressed genes whereas red cross (‘×’) indicates the deletion of genes in dark gray. Experimental data are presented as mean values with standard deviations from three independent experiments. Source data are provided as a Source Data file.