Abstract Astaxanthin exhibiting strong antioxidant activity, is widely used as food and animal feed additives as well as in cosmetics, pharmaceutical and nutraceuticals industries. Microbial production of astaxanthin has received increasing concerns in recent years. A combined strategy consisting of physical mutagenesis by atmospheric and room temperature plasma (ARTP) and adaptive evolution driven by H2O2, was established to expand disturbance in genome of the engineered Saccharomyces cerevisiae. A titer of 65.9 mg/L astaxanthin at flask level was obtained via multiple rounds of crossed ARTP and H2O2 treatment, which was increased nearly 4-fold compared with the starting strain. Moreover, a trend of decreasing intracellular reactive oxygen species (ROS) and increasing chronological lifespan (CLS) was observed in cells with improved titer of astaxanthin. Eventually, a highest reported titer of 404.78 mg/L astaxanthin in S. cerevisiae was achieved in 5-L fermenter by further optimization of fermentation conditions. Our study not only highlights the iterative cycling of the integrated strategy combined by ARTP and adaptive evolution tightly based on the natural properties of the targets provides an efficient route to boost product accumulation, but also demonstrates it can serve as a general strategy for other microbial cell factories.

中文翻译：

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通过结合诱变和进化提高酵母中虾青素的产量

摘要 虾青素具有很强的抗氧化活性，被广泛用作食品和动物饲料添加剂以及化妆品、制药和保健品行业。近年来，虾青素的微生物生产受到越来越多的关注。建立了由大气和室温等离子体 (ARTP) 物理诱变和 H2O2 驱动的适应性进化组成的组合策略，以扩大工程酿酒酵母基因组的干扰。通过多轮交叉ARTP和H2O2处理获得了65.9mg/L的烧瓶虾青素滴度，与起始菌株相比增加了近4倍。而且，在虾青素滴度提高的细胞中观察到细胞内活性氧 (ROS) 减少和实足寿命 (CLS) 增加的趋势。最终，通过进一步优化发酵条件，在 5-L 发酵罐中实现了酿酒酵母中 404.78 mg/L 虾青素的最高报告效价。我们的研究不仅强调了 ARTP 与紧密基于目标自然特性的适应性进化相结合的集成策略的迭代循环，提供了一条促进产品积累的有效途径，而且还表明它可以作为其他微生物细胞的通用策略。工厂。