5-Methyluridine (5-MU) is a prominent intermediate for industrial synthesis of several antiviral-drugs, however, its availability over the past decades has overwhelmingly relied on chemical and enzymatic strategies. Here, we have realized efficient production of 5-MU in E. coli, for the first time, via a designer artificial pathway consisting of a two-enzyme cascade (UMP 5-methylase and phosphatase). More importantly, we have engineered the E. coli cell factory to boost 5-MU production by systematic evaluation of multiple strategies, and as a proof of concept, we have further developed an antibiotic-free fermentation strategy to realize 5-MU production (10.71 g/L) in E. coli MB229 (a ΔthyA strain). Remarkably, we have also established a versatile and robust platform with exploitation of the engineered E. coli for efficient production of diversified UMP-derived chemicals. This study paves the way for future engineering of E. coli as a synthetic biology platform for acceleratively accessing UMP-derived chemical diversities.

中文翻译：

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开发大肠杆菌平台以高效生产 UMP 衍生化学品


5-甲基尿苷 （5-MU） 是几种抗病毒药物工业合成的重要中间体，然而，在过去几十年中，它的可用性在很大程度上依赖于化学和酶策略。在这里，我们首次通过由两种酶级联反应（UMP 5-甲基化酶和磷酸酶）组成的设计人工途径在大肠杆菌中实现了 5-MU 的高效生产。更重要的是，我们设计了大肠杆菌细胞工厂，通过系统评估多种策略来提高 5-MU 的产生，作为概念验证，我们进一步开发了一种无抗生素发酵策略，以实现在大肠杆菌 MB229（一种 ΔthyA 菌株）中产生 5-MU （10.71 g/L）。值得注意的是，我们还建立了一个多功能且强大的平台，利用工程化大肠杆菌高效生产多样化的 UMP 衍生化学品。这项研究为未来将大肠杆菌工程化为加速访问 UMP 衍生的化学多样性的合成生物学平台铺平了道路。