Fig. 5

Shikimate pathway refactoring for enhanced 2-PE biosynthesis. a Schematic representation of pathway constructs for the library consisting of all 27 combinations of P_KmTEF3, P_KmPGK, and P_KmTDH3 promoters for expression of KmARO4^K221L, KmARO7^G141S, and KmPHA2. b OD₆₀₀, extracellular accumulation of 2-phenylethanol (2-PE), 2-phenylethyl acetate (2-PEAc) and phenylpyruvate (PP) in all refactored pathway combinations after 72 h of cultivation at 30, 37 and 45 °C in 25 mL YPD medium. The negative control strain (Cntrl.) is K. marxianus CBS 6556 ura3Δ his3Δ. c Extracellular formation of 2-PE, 2-PEAc and PP over time and at different temperatures for K. marxianus CBS 6556 ura3Δ his3Δ with refactored Shikimate pathway combination 4 (Comb.4). All experiments were performed in biological triplicates. Bars represent the mean, while error bars represent the standard deviation