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| Main Authors: | , , , , , , , , , , , , |
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| Format: | Artículo Open Access |
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Wiley
2024
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| Online Access: | https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bit.28721 |
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| _version_ | 1867006063787638784 |
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| author | Chong Qiu Xiaoge Wang Jiaojiao Zuo Runyang Li Cong Gao Xiulai Chen Jia Liu Wanqing Wei Jing Wu Guipeng Hu Wei Song Nan Xu Liming Liu |
| author_facet | Chong Qiu Xiaoge Wang Jiaojiao Zuo Runyang Li Cong Gao Xiulai Chen Jia Liu Wanqing Wei Jing Wu Guipeng Hu Wei Song Nan Xu Liming Liu Chong Qiu Xiaoge Wang Jiaojiao Zuo Runyang Li Cong Gao Xiulai Chen Jia Liu Wanqing Wei Jing Wu Guipeng Hu Wei Song Nan Xu Liming Liu |
| collection | Wiley Open Access |
| contents | Systems engineering Escherichia coli for efficient production p‐coumaric acid from glucose Chong Qiu Xiaoge Wang Jiaojiao Zuo Runyang Li Cong Gao Xiulai Chen Jia Liu Wanqing Wei Jing Wu Guipeng Hu Wei Song Nan Xu Liming Liu Biotechnology and Bioengineering AbstractP‐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. 10.1002/bit.28721 http://onlinelibrary.wiley.com/termsAndConditions#vor |
| doi_str_mv | 10.1002/bit.28721 |
| format | Artículo Open Access |
| id | wiley_oa_10_1002_bit_28721 |
| institution | Wiley Open Access |
| license_str_mv | http://onlinelibrary.wiley.com/termsAndConditions#vor |
| publishDate | 2024 |
| publisher | Wiley |
| record_format | wiley_oa |
| spellingShingle | Systems engineering Escherichia coli for efficient production p‐coumaric acid from glucose Chong Qiu Xiaoge Wang Jiaojiao Zuo Runyang Li Cong Gao Xiulai Chen Jia Liu Wanqing Wei Jing Wu Guipeng Hu Wei Song Nan Xu Liming Liu Biotechnology and Bioengineering Systems engineering Escherichia coli for efficient production p‐coumaric acid from glucose Chong Qiu Xiaoge Wang Jiaojiao Zuo Runyang Li Cong Gao Xiulai Chen Jia Liu Wanqing Wei Jing Wu Guipeng Hu Wei Song Nan Xu Liming Liu Biotechnology and Bioengineering AbstractP‐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. 10.1002/bit.28721 http://onlinelibrary.wiley.com/termsAndConditions#vor |
| title | Systems engineering Escherichia coli for efficient production p‐coumaric acid from glucose |
| topic | Biotechnology and Bioengineering |
| url | https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bit.28721 |