大肠杆菌
生物传感器
高通量筛选
基因工程
合成生物学
蛋白质工程
吞吐量
定向进化
化学
计算生物学
纳米技术
生物化学
组合化学
生化工程
计算机科学
生物
材料科学
工程类
酶
基因
突变体
电信
无线
作者
Qi Wang,Minjun Jia,Hongjie Li,Qingbin Li,Jian Zhang,Tianyuan Su,Zhiyong Cui,Qingsheng Qi,Qian Wang
出处
期刊:ACS Sustainable Chemistry & Engineering
[American Chemical Society]
日期:2024-03-11
标识
DOI:10.1021/acssuschemeng.3c06991
摘要
5-Aminolevulinic acid (ALA) is a multifunctional nonprotein amino acid used in agriculture, medicine, and other fields. As current environmental problems become increasingly serious, it is of great significance to establish a cheap, environmentally friendly, and sustainable way to synthesize ALA. Therefore, the biosynthesis of ALA is receiving increasing attention. However, the lack of high-throughput screening methods for ALA has become a limiting factor in improving microbial production of ALA. In this study, a high-throughput screening method was developed based on the relationship between reactive oxygen species (ROS) caused by ALA and cyclic adenosine monophosphate (cAMP). First, the relationship between the ROS accumulation and changes in cAMP levels was verified. Subsequently, the selected promoter was optimized by adding cAMP receptor protein (CRP) binding sites at its upstream, and a high-throughput screening method for ALA was established. HemA mutant and ALA-producing Escherichia coli strain mutant were obtained and combined with a series of metabolic engineering strategies to improve ALA production. Finally, the ALA fed-batch fermentation in a 5 L fermenter achieved the highest ALA titer of 58.54 g/L with a productivity of 1.58 g/L/h. This laid the foundation for the industrialization of ALA through biosynthesis.
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