Integration Site Library for Efficient Construction of Plasmid-Free Microbial Cell Factories in Escherichia coli

IF 5.7 1区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Journal of Agricultural and Food Chemistry Pub Date : 2024-10-26 DOI:10.1021/acs.jafc.4c08290

Xiaolei Wang, Liangyu Lu, Qiyuan Liu, Jinyi Li, Tong Wang, Jia Wang, Xinxiao Sun, Xiaolin Shen, Qipeng Yuan

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Abstract

Enhanced production stability and efficiency along with a decrease in production costs are required to build efficient microbial cell factories. Target genes can be integrated into the genome to enhance genetic stability, reduce reliance on antibiotics, and alleviate the metabolic burden. However, selecting the optimal insertion site for the desired gene expression levels remains challenging. Therefore, 18 commonly usedEscherichia coliintegration sites were systematically characterized in this study. Promoters of different strengths were combined with integration sites, yielding a differential intensity range of up to 93-fold. This indicated the versatility and precision of this approach for controlling gene expression levels. Referring to the library, pathway genes were strategically integrated into theE. coligenome based on their respective expression levels. Genetically stable and highly efficient engineered strains that could biosynthesize arbutin and p-aminobenzoic acid were constructed.

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高效构建大肠杆菌无质粒微生物细胞工厂的整合位点库

要建立高效的微生物细胞工厂，就必须提高生产稳定性和效率，同时降低生产成本。目标基因可以整合到基因组中，以提高遗传稳定性，减少对抗生素的依赖，减轻代谢负担。然而，为达到理想的基因表达水平而选择最佳插入位点仍是一项挑战。因此，本研究对 18 个常用的大肠杆菌整合位点进行了系统鉴定。不同强度的启动子与整合位点相结合，产生了高达 93 倍的不同强度范围。这表明这种方法在控制基因表达水平方面具有多功能性和精确性。参照文库，根据各自的表达水平，将通路基因策略性地整合到大肠杆菌基因组中。构建出了能够生物合成熊果苷和对氨基苯甲酸的基因稳定、高效的工程菌株。

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来源期刊

Journal of Agricultural and Food Chemistry 农林科学-农业综合

CiteScore

9.90

自引率

8.20%

发文量

1375

审稿时长

2.3 months

期刊介绍： The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.