代谢工程
大肠杆菌
清脆的
基因敲除
基因
Cas9
代谢途径
基因组编辑
重组DNA
拉伤
生物化学
基因组工程
球形红杆菌
合理设计
生物
化学
计算生物学
细菌
遗传学
解剖
光合作用
作者
Changchuan Ye,Yuting Yang,Xi Chen,Lijie Yang,Xiao Hua,Mengjie Yang,Xiangfang Zeng,Shiyan Qiao
出处
期刊:Research Square - Research Square
日期:2022-07-19
标识
DOI:10.21203/rs.3.rs-1845109/v1
摘要
Abstract 5-Aminolevulinic acid (ALA) recently received much attention due to its potential application in many fields. In this study, an ALA production strain of Escherichia coli was constructed by rational metabolic engineering and stepwise improvement based on known regulatory and metabolic information and CRISPR/Cas9 mediated gene knockout. A metabolic strategy to produce ALA directly from glucose in this recombinant E. coli via the C5 pathway was applied herein. The rational metabolic engineering by gene knockouts significantly improved ALA production from 662.3 to 1601.7 mg/L. In addition, we tried to synergistically produce ALA via the C4 pathway in recombinant strain. The expression of a modified hemA gene, encoding an ALA synthase from Rhodobacter sphaeroides , improved ALA production from 1601.7 to 2099.7 mg/L. After 24 h cultivation, a yield of 0.210 g ALA per g glucose was achieved by constructed E. coli D5:FYABD. These results suggest that an industrially competitive strain can be efficiently developed by metabolic engineering based on combined rational modification and optimization of gene expression.
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