Qiu Zetian, Han Yumei, Li Jia, Ren Yi, Liu Xue, Li Shengying, Zhao Guang-Rong, Du Lei
State Key Laboratory of Synthetic Biology, School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Jinnan District, Tianjin, 300350, China; State Key Laboratory of Microbial Technology, Shandong University, Qingdao, Shandong, 266237, China; Georgia Tech Shenzhen Institute, Tianjin University, Dashi Road 1, Nanshan District, Shenzhen, 518055, China.
State Key Laboratory of Synthetic Biology, School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Jinnan District, Tianjin, 300350, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China; Georgia Tech Shenzhen Institute, Tianjin University, Dashi Road 1, Nanshan District, Shenzhen, 518055, China.
Metab Eng. 2025 May;89:60-75. doi: 10.1016/j.ymben.2025.02.001. Epub 2025 Feb 11.
Heterologous biosynthesis of natural products with long biosynthetic pathways in microorganisms often suffers from diverse problems, such as enzyme promiscuity and metabolic burden. Flavonoids and their glycosides are important phytochemicals in the diet of human beings, with various health benefits and biological activities. Despite previous efforts and achievements, efficient microbial production of plant-derived flavonoid compounds with long pathways remains challenging. Herein, we applied metabolic division engineering of Escherichia coli consortia to overcome these limitations. By establishing new biosynthetic pathways, rationally adjusting metabolic node intermediates, and engineering different auxotrophic and orthogonal carbon sources for hosts, we established stable two- and three-bacteria co-culture systems to efficiently de novo produce 12 flavonoids (61.15-325.31 mg/L) and 36 corresponding flavonoid glycosides (1.31-191.79 mg/L). Furthermore, the co-culture system was rapidly extended in a plug-and-play manner to produce isoflavonoids, dihydrochalcones, and their glycosides. This study successfully alleviates metabolic burden and overcomes enzyme promiscuity, and provides significant insights that could guide the biosynthesis of other complex secondary metabolites.
在微生物中对具有长生物合成途径的天然产物进行异源生物合成通常会遇到各种问题，例如酶的混杂性和代谢负担。黄酮类化合物及其糖苷是人类饮食中重要的植物化学物质，具有多种健康益处和生物活性。尽管此前已做出努力并取得了一些成果，但利用微生物高效生产具有长途径的植物源黄酮类化合物仍然具有挑战性。在此，我们应用大肠杆菌联合体的代谢分区工程来克服这些限制。通过建立新的生物合成途径、合理调整代谢节点中间体以及为宿主设计不同的营养缺陷型和正交碳源，我们建立了稳定的双菌和三菌共培养系统，以高效地从头生产12种黄酮类化合物（61.15 - 325.31毫克/升）和36种相应的黄酮类糖苷（1.31 - 191.79毫克/升）。此外，该共培养系统以即插即用的方式迅速扩展，用于生产异黄酮、二氢查耳酮及其糖苷。本研究成功减轻了代谢负担并克服了酶的混杂性，并为指导其他复杂次生代谢物的生物合成提供了重要见解。
