Enhanced astaxanthin production in yeast via combined mutagenesis and evolution
Enhanced astaxanthin production in yeast via combined mutagenesis and evolution
复制标题
通过组合诱变和进化提高酵母中虾青素的产量
DOI：
10.1016/j.bej.2020.107519
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发表时间：
2020-04-15
影响因子：
3.9
通讯作者：
Yuan, Yingjin
中科院分区：
文献类型：
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作者：
Jiang, Guozhen;Yang, Zuming;Yuan, Yingjin
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、制药和营养品工业。近年来，微生物法生产虾青素受到越来越多的关注。采用大气压和室温等离子体物理诱变（ARTP）和过氧化氢（H2 O2）驱动的适应性进化相结合的策略，对酿酒酵母基因组进行了干扰扩展。通过多轮ARTP和H2 O2的交叉处理，在摇瓶水平上获得了65.9mg/L的虾青素效价，与起始菌株相比提高了近4倍。此外，在具有提高的虾青素滴度的细胞中观察到细胞内活性氧（ROS）减少和时间寿命（CLS）增加的趋势。最终，在S.通过进一步优化发酵条件，在5L发酵罐中获得了高产菌株。我们的研究不仅突出了ARTP和适应性进化紧密结合的集成策略的迭代循环，提供了一种有效的途径来促进产品积累，而且还表明它可以作为其他微生物细胞工厂的通用策略。
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.