Adaptive Evolution\nand Metabolic Engineering Boost\nLycopene Production in Saccharomyces cerevisiae via Enhanced Precursors Supply and Utilization

Lycopene is a red carotenoid with\nremarkable antioxidant\nactivity,\nwhich has been widely used in food, cosmetics, medicine, and other\nindustries. Production of lycopene in Saccharomyces\ncerevisiae provides an economic and sustainable means.\nMany efforts have been done in recent years, but the titer of lycopene\nseems to reach a ceiling. Enhancing the supply and utilization of\nfarnesyl diphosphate (FPP) is generally regarded as an efficient strategy\nfor terpenoid production. Herein, an integrated strategy by means\nof atmospheric and room-temperature plasma (ARTP) mutagenesis combined\nwith H₂O₂-induced adaptive laboratory evolution\n(ALE) was proposed to improve the supply of upstream metabolic flux\ntoward FPP. Enhancing the expression of <i>CrtE</i> and\nintroducing an engineered <i>CrtI</i> mutant (Y160F&N576S)\nincreased the utilization of FPP toward lycopene. Consequently, the\ntiter of lycopene in the strain harboring the <i>Ura3</i> marker was increased by 60% to 703 mg/L (89.3 mg/g DCW) at the shake-flask\nlevel. Eventually, the highest reported titer of 8.15 g/L of lycopene\nin S. cerevisiae was achieved in a\n7 L bioreactor. The study highlights an effective strategy that the\nsynergistic complementarity of metabolic engineering and adaptive\nevolution facilitates natural product synthesis.