Multiple metabolic engineering of Saccharomyces cerevisiae for the production of lycopene

Jiaheng Liu, Minxia Song, Xianhao Xu, Yaokang Wu, Yanfeng Liu, Guocheng Du, Jianghua Li, Long Liu, Xueqin Lv
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Abstract

Lycopene is a high-value-added tetraterpenoid, which is widely used in cosmetics, medicine, food, and dietary supplements. The intracellular mevalonate pathway of Saccharomyces cerevisiae provides natural precursors for terpenoid product synthesis, so it is an excellent host for the heterologous production of lycopene. In this study, a recombinant strain named L10 with efficient lycopene production capability was constructed through multiple strategies, such as regulating the gene copy number of key enzymes, increasing nicotinamide adenine dinucleotide phosphate supply, and reducing squalene accumulation. Then, considering that intracellular lycopene accumulation can cause cytotoxicity to S. cerevisiae, we attempted to identify a transporter that can efficiently transport lycopene from intracellular to extracellular space. Molecular docking simulations predicted that the ATP-binding cassette transporter Snq2p may be a potential transporter of lycopene, and its function in promoting lycopene secretion was further determined by overexpression verification. The lycopene secretion titer of the strain L10Z2 overexpressing Snq2p increased to 16.5 times that of the control at the shake-flask level. After optimizing the galactose regulation system, the intracellular and secreted lycopene production of L11Z2 reached 2113.78 and 26.28 mg/L, respectively, after 150 h fed-batch culture in a 3-L bioreactor. This work provides a new research direction for efficient lycopene synthesis in S. cerevisiae cell factory.

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