De novo biosynthesis of berberine and halogenated benzylisoquinoline alkaloids in Saccharomyces cerevisiae.
De novo biosynthesis of berberine and halogenated benzylisoquinoline alkaloids in Saccharomyces cerevisiae.
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DOI：
10.1038/s42004-023-00821-9
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发表时间：
2023-02-09
影响因子：
5.9
通讯作者：
Li, Sijin
中科院分区：
文献类型：
--
作者：
Han, Jianing;Li, Sijin
黄连素是一种用途广泛的从植物中提取的苄基异喹啉生物碱。微生物制造已经成为一种很有前途的方法，可以找到有价值的偏见。在这里，我们通过设计19个基因，其中包括12个来自植物和细菌的异源基因，证明了黄连素在酿酒酵母中的完全生物合成。过量表达瓶颈酶、发酵放大和发酵后加热处理使黄连素效价提高了643倍，达到1.08 mg L~(-1)。这条途径还显示了掺入卤化酪氨酸合成具有较高治疗潜力的非天然BIA衍生物的高效率。我们首次证明了11-氟-四氢柱胺的生物合成是通过九个酶反应实现的。我们的途径的效率和混杂性也允许两个氟取代的酪氨酸衍生物同时掺入8，3‘-二氟-椰子碱。这项工作突出了酵母作为多功能微生物生物合成平台在加强现有药物供应链和促进药物开发方面的潜力。黄连素是一种从植物中提取的具有多种药理活性的苄基异喹啉生物碱，然而，从药用植物和有机合成中生产小檗碱仍然是不可持续的。在这里，作者报道了通过改造来自植物和细菌的基因，在酵母中完全生物合成黄连素。
Berberine is an extensively used pharmaceutical benzylisoquinoline alkaloid (BIA) derived from plants. Microbial manufacturing has emerged as a promising approach to source valuable BIAs. Here, we demonstrated the complete biosynthesis of berberine in Saccharomyces cerevisiae by engineering 19 genes including 12 heterologous genes from plants and bacteria. Overexpressing bottleneck enzymes, fermentation scale-up, and heating treatment after fermentation increased berberine titer by 643-fold to 1.08 mg L-1. This pathway also showed high efficiency to incorporate halogenated tyrosine for the synthesis of unnatural BIA derivatives that have higher therapeutical potentials. We firstly demonstrate the in vivo biosynthesis of 11-fluoro-tetrahydrocolumbamine via nine enzymatic reactions. The efficiency and promiscuity of our pathway also allow for the simultaneous incorporation of two fluorine-substituted tyrosine derivatives to 8, 3’-di-fluoro-coclaurine. This work highlights the potential of yeast as a versatile microbial biosynthetic platform to strengthen current pharmaceutical supply chain and to advance drug development. Berberine is a plant-derived benzylisoquinoline alkaloid with diverse pharmaceutical activities, however, the production of berberine from medicinal plants and organic synthesis remains unsustainable. Here, the authors report the complete biosynthesis of berberine in yeast by engineering genes from plants and bacteria.
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影响因子：
4.8
作者：
Hagel, Jillian M.;Beaudoin, Guillaume A. W.;Facchini, Peter J.
通讯作者：
Facchini, Peter J.
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
影响因子：
4.3
作者：
Gesell, Andreas;Diaz Chavez, Maria Luisa;Kutchan, Toni M.
通讯作者：
Kutchan, Toni M.
DOI：
10.1073/pnas.2205848119
发表时间：
2022-08-16
影响因子：
11.1
作者：
通讯作者：
--
DOI：
10.1002/anie.201409164
发表时间：
2014-12-22
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Gatland AE;Pilgrim BS;Procopiou PA;Donohoe TJ
通讯作者：
Donohoe TJ