Enhanced Biosynthesis of Dihydromyricetin in Saccharomyces cerevisiae by Coexpression of Multiple Hydroxylases
Enhanced Biosynthesis of Dihydromyricetin in Saccharomyces cerevisiae by Coexpression of Multiple Hydroxylases
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通过共表达多种羟化酶增强酿酒酵母中二氢杨梅素的生物合成
DOI：
10.1021/acs.jafc.0c05261
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发表时间：
2020-12-02
影响因子：
6.1
通讯作者：
Zhou, Jingwen
中科院分区：
文献类型：
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作者：
Li, Guangjian;Li, Hongbiao;Zhou, Jingwen
二氢杨梅素(DHM)是一种传统的从植物中提取的黄酮类化合物，具有一定的保健作用。本研究旨在对DHM生物生产菌株进行代谢工程。两株BK-11和BQ-21与类黄酮类3-羟基酶(F3‘H)整合，或同时整合类黄酮类3’-羟基酶(F3‘H)。所获得的菌株表达了GmCPR和SLF3‘5’H酶，并利用INO 1p和TdH 1p启动子进一步提高了酿酒酵母中柚皮苷元的DHM产量。通过多拷贝整合，在5 L生物反应器中添加2.5g/L柚皮苷元，可获得709.6 mg/L二氢呋喃，这表明F3‘H和类黄酮3’S‘-羟基酶之间的协同作用可能促进了二氢呋喃的产生。通过删除分支途径基因，整合PhCHS、MsCHI、PC4CL和FjTAL，从葡萄糖中获得了246.4 mg/L DHM。据我们所知，这是报道的从头合成DHM的最高产量。
Dihydromyricetin (DHM) is a traditional plant-extracted flavonoid with some health benefits. This study aimed to metabolically engineer the strains for DHM bioproduction. Two strains of BK-11 and BQ-21 were integrated with flavonoid 3-hydroxylase (F3H) or both F3H and flavonoid 3'-hydroxylase (F3'H). The resulting strains have expressed the enzymes of GmCPR and SlF3'5'H, and then, the promoters of INO 1p and TDH 1p were used to enhance further the DHM production from naringenin in Saccharomyces cerevisiae. Through multiple-copy integration, 709.6 mg/L DHM was obtained by adding 2.5 g/L naringenin in a 5 L bioreactor, implying that the synergistic effect between F3'H and flavonoid 3'S'-hydroxylase is likely to promote the DHM production. An yield of 246.4 mg/L DHM was obtained from glucose by deleting genes for branch pathways and integrating PhCHS, MsCHI, Pc4CL, and FjTAL. To our knowledge, this is the highest production reported for the de novo biosynthesis of DHM.