Improved cryptic plasmids in probiotic Escherichia coli Nissle 1917 for antibiotic-free pathway engineering

被引:3
作者
Dong, Miao-Miao [1 ]
Song, Lu [1 ]
Xu, Jia-Qi [1 ]
Zhu, Lin [1 ]
Xiong, Liang-Bin [1 ,2 ]
Wei, Dong-Zhi [1 ]
Wang, Feng-Qing [1 ]
机构
[1] East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China
[2] Shanghai Univ Med & Hlth Sci, Sch Pharm, Shanghai 201318, Peoples R China
基金
上海市自然科学基金; 中国国家自然科学基金;
关键词
Escherichia coli Nissle 1917; Cryptic plasmid; Metabolic engineering; Salicylic acid; Ergothioneine; CELLS;
D O I
10.1007/s00253-023-12662-6
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The engineered probiotic Escherichia coli Nissle 1917 (EcN) is expected to be employed in the diagnosis and treatment of various diseases. However, the introduced plasmids typically require antibiotics to maintain genetic stability, and the cryptic plasmids in EcN are usually eliminated to avoid plasmid incompatibility which may change the inherent probiotic characteristics. Here, we provided a simple design to minimize the genetic change of probiotics by eliminating native plasmids and reintroducing the recombinants carrying functional genes. Specific insertion sites in the vectors showed significant differences in the expression of fluorescence proteins. Selected integration sites were applied in the de novo synthesis of salicylic acid, leading to a titer of 142.0 +/- 6.0 mg/L in a shake flask with good production stability. Additionally, the design successfully realized the biosynthesis of ergothioneine (45 mg/L) by one-step construction. This work expands the application scope of native cryptic plasmids to the easy construction of functional pathways.
引用
收藏
页码:5257 / 5267
页数:11
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