Abstract: As one of the important aromatic amino acids, L-phenylalanine has been widely used in food, chemical industry, medicine and other industrial products. In this study, the central metabolic pathway and shikimic acid pathway of Saccharomyces cerevisiae were rational reconstructed, and a preliminary enhanced strain of L-phenylalanine synthesis was obtained. The production of extracellular L-phenylalanine and L-tyrosine was 2.49 mg/L and 6.54 mg/L, respectively. In order to further enhance the accumulation of L-phenylalanine, ARO10 and PDC5 genes were knocked out. Furthermore, TYR1 was knocked out to block the competitive L-tyrosine synthesis pathway, and the extracellular L-phenylalanine production increased to 45.40 mg/L, which is the highest extracellular production for de novo synthesis of L-phenylalanine in S. cerevisiae.
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