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Systems engineering Escherichia coli for efficient production p-coumaric acid from glucose.
Qiu, Chong; Wang, Xiaoge; Zuo, Jiaojiao; Li, Runyang; Gao, Cong; Chen, Xiulai; Liu, Jia; Wei, Wanqing; Wu, Jing; Hu, Guipeng; Song, Wei; Xu, Nan; Liu, Liming.
Afiliação
  • Qiu C; College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, China.
  • Wang X; School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, China.
  • Zuo J; School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, China.
  • Li R; College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, China.
  • Gao C; College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, China.
  • Chen X; School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, China.
  • Liu J; School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, China.
  • Wei W; School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, China.
  • Wu J; School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi, China.
  • Hu G; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, China.
  • Song W; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, China.
  • Xu N; School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, China.
  • Liu L; College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, China.
Biotechnol Bioeng ; 121(7): 2147-2162, 2024 Jul.
Article em En | MEDLINE | ID: mdl-38666765
ABSTRACT
P-coumaric acid (p-CA), a pant metabolite with antioxidant and anti-inflammatory activity, is extensively utilized in biomedicine, food, and cosmetics industry. In this study, a synthetic pathway (PAL) for p-CA was designed, integrating three enzymes (AtPAL2, AtC4H, AtATR2) into a higher l-phenylalanine-producing strain Escherichia coli PHE05. However, the lower soluble expression and activity of AtC4H in the PAL pathway was a bottleneck for increasing p-CA titers. To overcome this limitation, the soluble expression of AtC4H was enhanced through N-terminal modifications. And an optimal mutant, AtC4HL373T/G211H, which exhibited a 4.3-fold higher kcat/Km value compared to the wild type, was developed. In addition, metabolic engineering strategies were employed to increase the intracellular NADPH pool. Overexpression of ppnk in engineered E. coli PHCA20 led to a 13.9-folds, 1.3-folds, and 29.1% in NADPH content, the NADPH/NADP+ ratio and p-CA titer, respectively. These optimizations significantly enhance p-CA production, in a 5-L fermenter using fed-batch fermentation, the p-CA titer, yield and productivity of engineered strain E. coli PHCA20 were 3.09 g/L, 20.01 mg/g glucose, and 49.05 mg/L/h, respectively. The results presented here provide a novel way to efficiently produce the plant metabolites using an industrial strain.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Propionatos / Ácidos Cumáricos / Escherichia coli / Engenharia Metabólica / Glucose Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Propionatos / Ácidos Cumáricos / Escherichia coli / Engenharia Metabólica / Glucose Idioma: En Ano de publicação: 2024 Tipo de documento: Article