当前位置: X-MOL 学术Metab. Eng. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Design and construction of an artificial pathway for biosynthesis of acetaminophen in Escherichia coli
Metabolic Engineering ( IF 8.4 ) Pub Date : 2021-09-03 , DOI: 10.1016/j.ymben.2021.09.001
Xiaolin Shen 1 , Xin Chen 1 , Jian Wang 2 , Xinxiao Sun 1 , Sifan Dong 1 , Yang Li 1 , Yajun Yan 2 , Jia Wang 1 , Qipeng Yuan 1
Affiliation  

Acetaminophen (AAP) is one of the most commonly used drug ingredients that possesses antipyretic and analgesic effects. As an unnatural chemical, AAP is commercially produced by chemical processes using petroleum-derived carbohydrates, such as phenol, as raw materials, which is unsustainable and eco-unfriendly. In this study, we report design and construction of an artificial biosynthetic pathway for de novo production of AAP from simple carbon source. By exploring and expanding the substrate repertoire of natural enzymes, we identified and characterized a novel p-aminobenzoic acid (p-ABA) monooxygenase and an p-aminophenol (p-AP) N-acetyltransferase, which enabled the bacterial production of AAP from p-ABA. Then, we constructed an p-ABA over-producer by screening of p-ABA synthases and enhancing glutamine availability, resulting in 836.43 mg/L p-ABA in shake flasks in E. coli. Subsequent assembly of the entire biosynthetic pathway permitted the de novo production of AAP from glycerol for the first time. Finally, pathway engineering by dynamically regulating the expression of pathway genes via a temperature-inducible controller enabled production enhancement of AAP with a titer of 120.03 mg/L. This work not only constructs a microbial platform for AAP production, but also demonstrates design and construction of artificial biosynthetic pathways via discovering novel bioreactions based on existing enzymes.



中文翻译:

大肠杆菌生物合成对乙酰氨基酚人工途径的设计与构建

对乙酰氨基酚(AAP)是最常用的药物成分之一,具有解热和镇痛作用。作为一种非天然化学品,AAP 是通过使用石油衍生的碳水化合物(如苯酚)作为原材料的化学过程进行商业生产的,这是不可持续和不环保的。在这项研究中,我们报告了从简单碳源从头生产 AAP的人工生物合成途径的设计和构建。通过探索和扩展天然酶的底物库,我们鉴定并表征了一种新型氨基苯甲酸 ( p- ABA) 单加氧酶和一种氨基苯酚 ( p- AP) N-乙酰转移酶,使细菌能够从p -ABA产生 AAP 。然后,我们通过筛选p -ABA 合酶和提高谷氨酰胺可用性构建了p -ABA 过量生产者,在大肠杆菌的摇瓶中产生了 836.43 mg/L 的p -ABA 。整个生物合成途径的后续组装允许从头首次从甘油生产 AAP。最后,通过温度诱导控制器动态调节通路基因表达的通路工程使 AAP 的产量提高,滴度为 120.03 mg/L。这项工作不仅构建了用于 AAP 生产的微生物平台,而且还通过发现基于现有酶的新型生物反应来展示人工生物合成途径的设计和构建。

更新日期:2021-09-06
down
wechat
bug