Violaxanthin is a plant-derived orange xanthophyll with remarkable antioxidant activity that has wide applications in various industries, such as food, agriculture, and cosmetics. In addition, it is the key precursor of important substances such as abscisic acid and fucoxanthin. Saccharomyces cerevisiae, as a GRAS (generally regarded as safe) chassis, provides a good platform for producing violaxanthin production with a yield of 7.3 mg/g DCW, which is far away from commercialization. Herein, an integrated strategy involving zeaxanthin epoxidase (ZEP) source screening, cytosol redox state engineering, and nicotinamide adenine dinucleotide phosphate (NADPH) regeneration was implemented to enhance violaxanthin production in S. cerevisiae. 58aa-truncated ZEP from Vitis vinifera exhibited optimal efficiency in an efficient zeaxanthin-producing strain. The titer of violaxanthin gradually increased by 17.9-fold (up to 119.2 mg/L, 15.19 mg/g DCW) via cytosol redox state engineering and NADPH supplementation. Furthermore, balancing redox homeostasis considerably improved the zeaxanthin concentration by 139.3% (up to 143.9 mg/L, 22.06 mg/g DCW). Thus, the highest reported titers of violaxanthin and zeaxanthin in S. cerevisiae were eventually achieved. This study not only builds an efficient platform for violaxanthin biosynthesis but also serves as a useful reference for the microbial production of xanthophylls.

中文翻译：

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酵母高效生产紫黄质的系统代谢工程

紫黄质是一种源自植物的橙色叶黄素，具有显着的抗氧化活性，在食品、农业、化妆品等各个行业有着广泛的应用。此外，它还是脱落酸和岩藻黄质等重要物质的关键前体。酿酒酵母作为GRAS（通常被认为是安全的）底物，为生产紫黄素提供了良好的平台，产量为7.3 mg/g DCW，距离商业化还很远。在此，实施了涉及玉米黄质环氧化酶（ZEP）来源筛选、细胞质氧化还原态工程和烟酰胺腺嘌呤二核苷酸磷酸（NADPH）再生的综合策略，以提高酿酒酵母中紫黄质的产量。来自葡萄的58aa 截短ZEP在高效玉米黄质生产菌株中表现出最佳效率。通过胞质氧化还原状态工程和NADPH补充，紫黄质的滴度逐渐增加了17.9倍（高达119.2 mg/L，15.19 mg/g DCW） 。此外，平衡氧化还原稳态显着提高了玉米黄质浓度 139.3%（高达 143.9 mg/L，22.06 mg/g DCW）。因此，最终实现了酿酒酵母中紫黄质和玉米黄质的最高报道滴度。该研究不仅构建了紫黄质生物合成的高效平台，也为微生物生产叶黄素提供了有益的参考。