Fig. 5: Engineering P. occidentalis Y-7552 for efficient synthesis of CCM.

a Heterologous pathway for synthesis of CCM from endogenous 3-dehydroshikimate (DHS). The native yeast shikimate pathway is shown in blue, while the heterologous CCM synthesis pathway is shaded and shown in red. b Titers of CCM pathway intermediates in culture supernatants of successive engineered P. occidentalis production strains. Metabolite titers are depicted in g L⁻¹ for PCA, CAT, and CCM. Asterisks (*) denote a significant increase or decrease in metabolite titer relative to the respective parent strain (P < 0.05). Statistical differences between parent and derivative strains were tested using two-tailed Student’s t-test. Error bars represent the mean ± s.d. of n = 3 independent biological samples. c Cultivation of a CCM-producing P. occidentalis strain (LP635) in a fed-batch fermentor using a mineral medium at pH 6.0. Growth of biomass (OD₆₀₀) and accumulation of CCM pathway metabolites in the culture medium were measured during cultivation. OD₆₀₀ is reported in arbitrary units (arb. units). CAT catechol, CCM cis,cis-muconic acid, DAHP 3-deoxy-d-arabinoheptulosonate 7-phosphate, DHQ 3-dehydroquinate, DHS 3-dehydroshikimate, E4P erythrose 4-phosphate, FMN flavin mononucleotide, PCA protocatechuic acid, PEP phosphoenolpyruvate, prFMN prenylated flavin mononucleotide. Source data are provided as a Source Data file.