Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coli
Abstract Acetic acid, a by-product of cytidine synthesis, competes for carbon flux from central metabolism, which may be directed either to the tricarboxylic acid (TCA) cycle for cytidine synthesis or to overflow metabolites, such as acetic acid. In Escherichia coli, the acetic acid synthesis pathwa...
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| Language: | English |
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BMC
2025-02-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-025-02657-5 |
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| author | Tong Ye Wei Ding Zhengxu An Haojie Zhang Xiaobo Wei Junnan Xu Huiyan Liu Haitian Fang |
| author_facet | Tong Ye Wei Ding Zhengxu An Haojie Zhang Xiaobo Wei Junnan Xu Huiyan Liu Haitian Fang |
| author_sort | Tong Ye |
| collection | DOAJ |
| description | Abstract Acetic acid, a by-product of cytidine synthesis, competes for carbon flux from central metabolism, which may be directed either to the tricarboxylic acid (TCA) cycle for cytidine synthesis or to overflow metabolites, such as acetic acid. In Escherichia coli, the acetic acid synthesis pathway, regulated by the poxB and pta genes, facilitates carbon consumption during cytidine production. To mitigate carbon source loss, the CRISPR-Cas9 gene-editing technique was employed to knock out the poxB and pta genes in E. coli, generating the engineered strains K12ΔpoxB and K12ΔpoxBΔpta. After 39 h of fermentation in 500 mL shake flasks, the cytidine yields of strains K12ΔpoxB and K12ΔpoxBΔpta were 1.91 ± 0.04 g/L and 18.28 ± 0.22 g/L, respectively. Disruption of the poxB and pta genes resulted in reduced acetic acid production and glucose consumption. Transcriptomic and metabolomic analyses revealed that impairing the acetic acid metabolic pathway in E. coli effectively redirected carbon flux toward cytidine biosynthesis, yielding a 5.26-fold reduction in acetate metabolism and an 11.56-fold increase in cytidine production. These findings provide novel insights into the influence of the acetate metabolic pathway on cytidine biosynthesis in E. coli. Graphical Abstract |
| format | Article |
| id | doaj-art-2f63229d95a640a4857082e4897b2dcf |
| institution | Kabale University |
| issn | 1475-2859 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj-art-2f63229d95a640a4857082e4897b2dcf2025-02-09T13:00:50ZengBMCMicrobial Cell Factories1475-28592025-02-0124111510.1186/s12934-025-02657-5Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coliTong Ye0Wei Ding1Zhengxu An2Haojie Zhang3Xiaobo Wei4Junnan Xu5Huiyan Liu6Haitian Fang7School of Life Sciences, Ningxia UniversitySchool of Food Science and Engineering, Ningxia UniversitySchool of Food Science and Engineering, Ningxia UniversitySchool of Food Science and Engineering, Ningxia UniversitySchool of Food Science and Engineering, Ningxia UniversitySchool of Food Science and Engineering, Ningxia UniversitySchool of Food Science and Engineering, Ningxia UniversitySchool of Life Sciences, Ningxia UniversityAbstract Acetic acid, a by-product of cytidine synthesis, competes for carbon flux from central metabolism, which may be directed either to the tricarboxylic acid (TCA) cycle for cytidine synthesis or to overflow metabolites, such as acetic acid. In Escherichia coli, the acetic acid synthesis pathway, regulated by the poxB and pta genes, facilitates carbon consumption during cytidine production. To mitigate carbon source loss, the CRISPR-Cas9 gene-editing technique was employed to knock out the poxB and pta genes in E. coli, generating the engineered strains K12ΔpoxB and K12ΔpoxBΔpta. After 39 h of fermentation in 500 mL shake flasks, the cytidine yields of strains K12ΔpoxB and K12ΔpoxBΔpta were 1.91 ± 0.04 g/L and 18.28 ± 0.22 g/L, respectively. Disruption of the poxB and pta genes resulted in reduced acetic acid production and glucose consumption. Transcriptomic and metabolomic analyses revealed that impairing the acetic acid metabolic pathway in E. coli effectively redirected carbon flux toward cytidine biosynthesis, yielding a 5.26-fold reduction in acetate metabolism and an 11.56-fold increase in cytidine production. These findings provide novel insights into the influence of the acetate metabolic pathway on cytidine biosynthesis in E. coli. Graphical Abstracthttps://doi.org/10.1186/s12934-025-02657-5Acetate metabolismCarbon metabolic fluxCytidineEscherichia coli |
| spellingShingle | Tong Ye Wei Ding Zhengxu An Haojie Zhang Xiaobo Wei Junnan Xu Huiyan Liu Haitian Fang Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coli Microbial Cell Factories Acetate metabolism Carbon metabolic flux Cytidine Escherichia coli |
| title | Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coli |
| title_full | Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coli |
| title_fullStr | Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coli |
| title_full_unstemmed | Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coli |
| title_short | Increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in Escherichia coli |
| title_sort | increased distribution of carbon metabolic flux during de novo cytidine biosynthesis via attenuation of the acetic acid metabolism pathway in escherichia coli |
| topic | Acetate metabolism Carbon metabolic flux Cytidine Escherichia coli |
| url | https://doi.org/10.1186/s12934-025-02657-5 |
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