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Metabolic engineering of a robust Escherichia coli strain with a dual protection system.
Ou, Xiao-Yang; Wu, Xiao-Ling; Peng, Fei; Zeng, Ying-Jie; Li, Hui-Xian; Xu, Pei; Chen, Gu; Guo, Ze-Wang; Yang, Ji-Guo; Zong, Min-Hua; Lou, Wen-Yong.
Afiliação
  • Ou XY; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Wu XL; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Peng F; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Zeng YJ; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Li HX; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Xu P; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Chen G; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Guo ZW; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Yang JG; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
  • Zong MH; Innovation Center of Bioactive Molecule Development and Application, South China Institute of Collaborative Innovation, Dongguan, China.
  • Lou WY; Lab of Applied Biocatalysis, School of Food Science and Engineering, South China University of Technology, Guangzhou, China.
Biotechnol Bioeng ; 116(12): 3333-3348, 2019 12.
Article em En | MEDLINE | ID: mdl-31502661
Considerable attention has been given to the development of robust fermentation processes, but microbial contamination and phage infection remain deadly threats that need to be addressed. In this study, a robust Escherichia coli BL21(DE3) strain was successfully constructed by simultaneously introducing a nitrogen and phosphorus (N&P) system in combination with a CRISPR/Cas9 system. The N&P metabolic pathways were able to express formamidase and phosphite dehydrogenase in the host cell, thus enabled cell growth in auxotrophic 3-(N-morpholino)propanesulfonic acid medium with formamide and phosphite as nitrogen and phosphorus sources, respectively. N&P metabolic pathways also allowed efficient expression of heterologous proteins, such as green fluorescent protein (GFP) and chitinase, while contaminating bacteria or yeast species could hardly survive in this medium. The host strain was further engineered by exploiting the CRISPR/Cas9 system to enhance the resistance against phage attack. The resultant strain was able to grow in the presence of T7 phage at a concentration of up to 2 × 107 plaque-forming units/ml and produce GFP with a yield of up to 30 µg/109 colony-forming units, exhibiting significant advantages over conventional engineered E. coli. This newly engineered, robust E. coli BL21(DE3) strain therefore shows great potential for future applications in industrial fermentation.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Bacteriófago T7 / Escherichia coli / Engenharia Metabólica / Microrganismos Geneticamente Modificados Idioma: En Ano de publicação: 2019 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Bacteriófago T7 / Escherichia coli / Engenharia Metabólica / Microrganismos Geneticamente Modificados Idioma: En Ano de publicação: 2019 Tipo de documento: Article