Metabolic Division in an Escherichia coli Coculture System for Efficient Production of Kaempferide
Metabolic Division in an Escherichia coli Coculture System for Efficient Production of Kaempferide
复制标题
大肠杆菌共培养系统中的代谢分裂以有效生产山柰苷
DOI：
10.1021/acssynbio.1c00510
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发表时间：
2022-03-18
影响因子：
4.7
通讯作者：
Zhao, Guang-Rong
中科院分区：
文献类型：
--
作者：
Qiu, Zetian;Liu, Xue;Zhao, Guang-Rong
山奈素是一种植物源天然黄酮类化合物，具有良好的药理活性，在人体保健中具有重要的药用和营养价值。复杂黄酮类化合物的高效微生物生产受到代谢串扰和代谢负担的影响，这是合成生物学面临的一个巨大挑战。在此，我们确定了4 '-O-甲基转移酶，并将山奈素的人工生物合成途径分为上游、中游和下游模块。通过组合来自不同来源的异源基因并微调表达，我们优化了用于生产山奈素的每个模块。此外，我们设计和评估的共培养系统中的合成劳动的四种分工模式，即插即用的模块化。三株共培养的三个模块的线性划分显示出比两株共培养更高的山柰素生产率。结果表明，采用“U”形分割，将上下游组件共分为一个菌株，使共培养系统的性能达到最佳，其山奈素产量为116.0 ± 3.9 mg/L，分别比单菌、双菌和三菌共培养线性分割的山奈素产量高510、140和50%.这是第一个报告的有效从头生产山柰素在一个强大的大肠杆菌共培养。共培养中的U形途径分裂策略为提高有价值的复杂天然产物的生产力提供了一条有前途的途径。
Kaempferide, a plant-derived natural flavonoid, exhibits excellent pharmacological activities with nutraceutical and medicinal applications in human healthcare. Efficient microbial production of complex flavonoids suffers from metabolic crosstalk and burden, which is a big challenge for synthetic biology. Herein, we identified 4'-O-methyltransferases and divided the artificial biosynthetic pathway of kaempferide into upstream, midstream, and downstream modules. By combining heterologous genes from different sources and fine-tuning the expression, we optimized each module for the production of kaempferide. Furthermore, we designed and evaluated four division patterns of synthetic labor in coculture systems by plug-and-play modularity. The linear division of three modules in a three-strain coculture showed higher productivity of kaempferide than that in two-strain cocultures. The U-shaped division by co-distributing the upstream and downstream modules in one strain led to the best performance of the coculture system, which produced 116.0 +/- 3.9 mg/L kaempferide, which was 510, 140, and 50% higher than that produced by the monoculture, two-strain coculture, and three-strain coculture with the linear division, respectively. This is the first report of efficient de novo production of kaempferide in a robust Escherichia coil coculture. The strategy of U-shaped pathway division in the coculture provides a promising way for improving the productivity of valuable and complex natural products.