Improve the Biosynthesis of Baicalein and Scutellarein via Manufacturing Self-Assembly Enzyme Reactor In Vivo
Improve the Biosynthesis of Baicalein and Scutellarein via Manufacturing Self-Assembly Enzyme Reactor In Vivo
复制标题
通过体内制造自组装酶反应器提高黄芩素和黄芩素的生物合成。
DOI：
10.1021/acssynbio.0c00606
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发表时间：
2021-04-21
影响因子：
4.7
通讯作者：
Wang, Yong
中科院分区：
文献类型：
--
作者：
Ji, Dongni;Li, Jianhua;Wang, Yong
黄芩素和黄芩素是从中药黄芩中提取的具有生物活性的黄酮类化合物，但目前还缺乏有效的生产方法。在这项研究中，我们开发了一种顺序自组装酶反应器，涉及黄芩苷途径中的两种酶以及大肠杆菌中的一对蛋白质-肽相互作用。杆菌这些结构域使我们能够优化两个黄芩素生物合成酶的化学计量招募是一个酶复合物。该策略减少了中间体的积累并消除了途径瓶颈。在摇瓶发酵中，黄芩素的效价提高了6.6倍（从21.6 mg/L提高到143.5 mg/L），黄芩素的效价提高了1.4倍（从84.3 mg/L提高到120.4 mg/L）。首次实现了以葡萄糖为底物的黄芩素生物合成，并通过补料分批发酵，获得了214.1mg/L的黄芩素产量。本研究为黄芩素和野黄芩素的发酵工艺优化和规模化生产提供了新的思路。
Baicalein and scutellarein are bioactive flavonoids isolated from the traditional Chinese medicine Scutellaria baicalensis Georgi; however, there is a lack of effective strategies for producing baicalein and scutellarein. In this study, we developed a sequential self-assembly enzyme reactor involving two enzymes in the baicalein pathway with a pair of protein-peptide interactions in E. coli. These domains enabled us to optimize the stoichiometry of two baicalein biosynthetic enzymes recruited to be an enzymes complex. This strategy reduces the accumulation of intermediates and removes the pathway bottleneck. With this strategy, we successfully promoted the titer of baicalein by 6.6-fold (from 21.6 to 143.5 mg/L) and that of scutellarein by 1.4-fold (from 84.3 to 120.4 mg/L) in a flask fermentation, respectively. Furthermore, we first achieved the de novo biosynthesis of baicalein directly from glucose, and the strain was capable of producing 214.1 mg/L baicalein by fed-batch fermentation. This work provides novel insights for future optimization and large-scale fermentation of baicalein and scutellarein.