Model-based dynamic engineering of Escherichia coli for N-acetylglucosamine overproduction
Model-based dynamic engineering of Escherichia coli for N-acetylglucosamine overproduction
复制标题
基于模型的大肠杆菌 N-乙酰氨基葡萄糖过量生产动态工程
DOI：
10.1016/j.biotno.2022.02.001
复制
发表时间：
2022-02
期刊：
影响因子：
--
通讯作者：
Long Liu
中科院分区：
文献类型：
--
作者：
Jiangong Lu;Yaokang Wu;Chen Deng;Yanfeng Liu;Xueqin Lv;Jianghua Li;Guocheng Du;Long Liu
N-acetylglucosamine (GlcNAc), a glucosamine derivative, has a wide range of applications in pharmaceutical fields, and there is an increasing interest in the efficient production of GlcNAc genetic engineered bacteria. In this work,Escherichia coliATCC 25947 (DE3) strain was engineered by a model-based dynamic regulation strategy achieving GlcNAc overproduction. First, the GlcNAc synthetic pathway was introduced intoE. coli, and through flux balance analysis of the genome-scale metabolic network model, metabolic engineering strategies were generated to further increase GlcNAc yield. Knock-out of genespoxBandldhA,encoding pyruvate oxidase and lactate dehydrogenase, increased GlcNAc titer by 5.1%. Furthermore, knocking out N-acetylmuramic acid 6-phosphate etherase encoded bymurQand enhancing glutamine synthetase encoded byglnAgene further increased GlcNAc titer to 130.8 g/L. Analysis of metabolic flux balance showed that GlcNAc production maximization requires the strict dynamic restriction of the reactions catalyzed bypfkAandzwfto balance cell growth and product synthesis. Hence, a dynamic regulatory system was constructed by combining the CRISPRi (clustered regularly interspaced short palindromic repeats interference) system with the lactose operonlacIand the transcription factor pdhR, allowing the cell to respond to the concentration of pyruvate and IPTG to dynamically represspfkAandzwftranscription. Finally, the engineered bacteria with the dynamic regulatory system produced 143.8 g/L GlcNAc in a 30-L bioreactor in 55 h with a yield reaching 0.539 g/g glucose. Taken together, this work significantly enhanced the GlcNAc production ofE. coli.Moreover, it provides a systematic, effective, and universal way to improve the synthetic ability of other engineered strains.
登录
查看更多内容
影响因子：
4.4
作者：
Jiang, Yu;Chen, Biao;Yang, Sheng
通讯作者：
Yang, Sheng
影响因子：
4
作者：
Schöpping M;Gaspar P;Neves AR;Franzén CJ;Zeidan AA
通讯作者：
Zeidan AA
影响因子：
5.7
作者：
Isupov, MN;Obmolova, G;Teplyakov, A
通讯作者：
Teplyakov, A
影响因子：
8.4
作者：
Lee, Sang-Woo;Oh, Min-Kyu
通讯作者：
Oh, Min-Kyu
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL