Fig. 2: Optimizing (S)-NOR production in yeast.
a Biosynthetic pathway from glucose to generate (S)-NOR in yeast. Color scheme of pathway: green, overexpression of endogenous genes or mutants; orange, introduction of heterologous genes; purple arrows, plant-based tyrosine pathway; black dashed arrow, multiple steps; red, gene deletion. EcAROL, shikimate kinase; MtPDH1, prephenate dehydrogenase; AtPAT, prephenate aminotransferase; MtncADH, noncanonical arogenate dehydrogenase; PsAAAD*, aromatic amino acid decarboxylase mutant. PPP, pentose phosphate pathway; E4P, erythrose 4-phosphate; PEP, phosphoenolpyruvate; DAHP, 3-deoxy-D-arabino-heptulosonate 7-phosphate; SA, shikimic acid; S3P, shikimate 3-phosphate; EPSP, 5-enolpyruvylshikimate-3-phosphate; CHA, chorismate; PPA, prephenate; HPP, 4-hydroxyphenylpyruvate; 4-HPAA, 4-hydroxyphenylacetaldehyde; Tyrosol, 2-(4-Hydroxyphenyl) ethanol; 4-HPAC, 4-Hydroxyphenylacetic acid. (S)-NOR titers and final OD600 in engineered strains with b the removal of potential aldehyde reductases or dehydrogenases; c increasing more copies of CjNCS∆35; d introducing aromatic amino acid synthase and e plant-based tyrosine pathway. At least three biologically independent colonies were grown for 72 h in 20 mL minimal media with 20 g/L glucose. Augmenting the flux indicates overexpression of yeast native ARO1, ARO2, ARO3, and expression of EcAROL, MtPDH1, ARO4K229L and ARO7G141S. (S)-NOR pathway indicates the introduction of optimal CYP76AD5, DODC and CjNCS∆35. Significance was calculated using two-tailed t-test. Data are presented as mean ± standard deviations (n = 3 or 4 biologically independent samples). Source data are provided as a Source Data file.
