Academic Journal
Dynamic flux regulation for high-titer anthranilate production by plasmid-free, conditionally-auxotrophic strains of Pseudomonas putida
| العنوان: | Dynamic flux regulation for high-titer anthranilate production by plasmid-free, conditionally-auxotrophic strains of Pseudomonas putida |
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| المؤلفون: | Fernández-Cabezón, Lorena, Rosich i Bosch, Berta, Kozaeva, Ekaterina, Gurdo, Nicolás, Nikel, Pablo Iván |
| المصدر: | Fernández-Cabezón , L , Rosich i Bosch , B , Kozaeva , E , Gurdo , N & Nikel , P I 2022 , ' Dynamic flux regulation for high-titer anthranilate production by plasmid-free, conditionally-auxotrophic strains of Pseudomonas putida ' , Metabolic Engineering , vol. 73 , pp. 11-25 . https://doi.org/10.1016/j.ymben.2022.05.008 |
| سنة النشر: | 2022 |
| المجموعة: | Technical University of Denmark: DTU Orbit / Danmarks Tekniske Universitet |
| مصطلحات موضوعية: | Anthranilate, Dynamic regulation, Metabolic engineering, Pseudomonas putida, Quorum sensing, Shikimate pathway, Synthetic biology, Synthetic metabolism |
| الوصف: | Anthranilate, an intermediate of the shikimate pathway, is a high-value aromatic compound widely used as a precursor in the production of dyes, fragrances, plastics and pharmaceuticals. Traditional strategies adopted for microbial anthranilate production rely on the implementation of auxotrophic strains—which requires aromatic amino acids or complex additives to be supplemented in the culture medium, negatively impacting production costs. In this work, we engineered the soil bacterium Pseudomonas putida for high-titer, glucose-dependent anthranilate production by repurposing elements of the Esa quorum sensing (QS) system of Pantoea stewartii. The P esaS promoter mediated a self-regulated transcriptional response that effectively knocked-down the expression of the trpDC genes. Next, we harnessed the synthetic QS elements to engineer a growth-to-anthranilate production switch. The resulting plasmid-free P. putida strain produced the target compound at 3.8 ± 0.3 mM in shaken-flask cultures after 72 h—a titer >2-fold higher than anthranilate levels reported thus far. Our results highlight the value of dynamic flux regulation for the production of intermediate metabolites within highly-regulated routes (such as the shikimate pathway), thereby circumventing the need of expensive additives. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| Relation: | https://orbit.dtu.dk/en/publications/98776a79-35d2-47ea-b800-3884556a1bf6 |
| DOI: | 10.1016/j.ymben.2022.05.008 |
| الاتاحة: | https://orbit.dtu.dk/en/publications/98776a79-35d2-47ea-b800-3884556a1bf6 https://doi.org/10.1016/j.ymben.2022.05.008 https://backend.orbit.dtu.dk/ws/files/298036036/1_s2.0_S1096717622000726_main.pdf |
| Rights: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.F2454FD5 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1016/j.ymben.2022.05.008 |
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