生物生产
生物化学
大肠杆菌
生物合成
代谢途径
生物
ATP结合盒运输机
ATP合酶
硫链球菌素
代谢工程
发酵
运输机
酶
基因
核糖核酸
核糖体
作者
Wenqiu Wang,Yulong Xiang,Guobin Yin,Shan Hu,Jian Cheng,Jian Chen,Guocheng Du,Zhen Kang,Yang Wang
标识
DOI:10.1021/acs.jafc.4c00903
摘要
5-Aminolevulinic acid (5-ALA) plays a pivotal role in the biosynthesis of heme and chlorophyll and has garnered great attention for its agricultural applications. This study explores the multifaceted construction of 5-ALA microbial cell factories. Evolutionary analysis-guided screening identified a novel 5-ALA synthase from Sphingobium amiense as the best synthase. An sRNA library facilitated global gene screening that demonstrated that trpC and ilvA repression enhanced 5-ALA production by 74.3% and 102%, respectively. Subsequently, efflux of 5-ALA by the transporter Gdx increased 5-ALA biosynthesis by 25.7%. To mitigate oxidative toxicity, DNA-binding proteins from starved cells were employed, enhancing cell density and 5-ALA titer by 21.1 and 4.1%, respectively. Combining these strategies resulted in an Escherichia coli strain that produced 5-ALA to 1.51 g·L–1 in shake flask experiments and 6.19 g·L–1 through fed-batch fermentation. This study broadens the repertoire of available 5-ALA synthases and transporters and provides a new platform for optimizing 5-ALA bioproduction.
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