Dynamic control of the plasmid copy number maintained without antibiotics in Escherichia coli DOI Creative Commons
Geunyung Park, Jina Yang, Sang Woo Seo

и другие.

Journal of Biological Engineering, Год журнала: 2024, Номер 18(1)

Опубликована: Дек. 19, 2024

Abstract Background Manipulating the gene expression is key strategy to optimize metabolic flux. Not only transcription, translation, and post-translation level control, but also dynamic plasmid copy number (PCN) control has been studied. The PCN systems that have developed date are based on understanding of origin replication mechanisms, which limits their application specific origins requires use antibiotics for maintenance. In this study, we a system Escherichia coli maintained without antibiotics. This achieved by regulating transcription translation initiation factor IF-1 ( infA ), an essential encoded plasmid, while deleting it from plasmid-bearing host cell. Results When validated using GFP as reporter protein, our demonstrated 22-fold range in within CloDF13 origin. was employed determine optimal carrying cad gene, converts intermediate tricarboxylic acid cycle (TCA cycle) itaconic acid. By optimizing PCN, could achieve titer 3 g/L, 5.3-fold higher than strain. Conclusions Our offers identify genes competitive relationship with pathways crucial growth organism. approach can potentially be applied other bacterial hosts substituting sensing module or gene.

Язык: Английский

Metabolic engineering of Corynebacterium glutamicum for highly selective production of 5-hydroxyvaleric acid DOI

Yu Jung Sohn,

Hee Taek Kim, Min‐Soo Kang

и другие.

Metabolic Engineering, Год журнала: 2025, Номер unknown

Опубликована: Март 1, 2025

Язык: Английский

Процитировано

0
Low-biomass pyruvate production with engineered Vibrio natriegens is accompanied by parapyruvate formation DOI Creative Commons
Maurice Hädrich,

Clarissa Scheuchenegger,

Sören-Tobias Vital

и другие.

Microbial Cell Factories, Год журнала: 2025, Номер 24(1)

Опубликована: Март 28, 2025

Abstract Background Pyruvate is a precursor for various compounds in the chemical, drug, and food industries therefore an attractive target molecule microbial production processes. The fast-growing bacterium Vibrio natriegens excels with its specific substrate uptake rate as unconventional chassis industrial biotechnology. Here, we aim to exploit traits of V. pyruvate fermentations low biomass concentrations. Results We inactivated dehydrogenase complex Δ vnp12 , which harbors deletions prophage regions . resulting strain aceE was unable grow minimal medium glucose unless supplemented acetate. In shaking flasks, showed growth 1.16 ± 0.03 h − 1 produced 4.0 0.3 g Pyr L within 5 h. optimized parameters aerobic fermentation process applied constant maintenance feed 0.24 Ac resulted maximal concentration only 6.6 0.4 CDW yielded highly active resting cells (q S ) 3.5 0.2 Glc −1 41.0 1.8 volumetric productivity 4.1 Carbon balancing disclosed gap 30%, identified partly parapyruvate. Deletion ligK encoding HMG/CHA aldolase did not impact formation but plasmid-based overexpression negatively affected led 3-fold higher parapyruvate culture broth. Notably, also supernatants pyruvate-producing Corynebacterium glutamicum strain. Cell-free bioreactor experiments mimicking biological formation, pointing chemical reaction contributing synthesis. Conclusions engineered metabolically producing high at concentration. However, found that accompanied by well C. Parapyruvate seems be result conversion might supported biochemically reaction.

Язык: Английский

Процитировано

0
Vibrio natriegens: Application of a Fast-Growing Halophilic Bacterium DOI
Maurice Hädrich, Clarissa Schulze, Josef Hoff

и другие.

Advances in biochemical engineering, biotechnology, Год журнала: 2024, Номер unknown

Опубликована: Янв. 1, 2024

Язык: Английский

Процитировано

1
Dynamic control of the plasmid copy number maintained without antibiotics in Escherichia coli DOI Creative Commons
Geunyung Park, Jina Yang, Sang Woo Seo

и другие.

Journal of Biological Engineering, Год журнала: 2024, Номер 18(1)

Опубликована: Дек. 19, 2024

Abstract Background Manipulating the gene expression is key strategy to optimize metabolic flux. Not only transcription, translation, and post-translation level control, but also dynamic plasmid copy number (PCN) control has been studied. The PCN systems that have developed date are based on understanding of origin replication mechanisms, which limits their application specific origins requires use antibiotics for maintenance. In this study, we a system Escherichia coli maintained without antibiotics. This achieved by regulating transcription translation initiation factor IF-1 ( infA ), an essential encoded plasmid, while deleting it from plasmid-bearing host cell. Results When validated using GFP as reporter protein, our demonstrated 22-fold range in within CloDF13 origin. was employed determine optimal carrying cad gene, converts intermediate tricarboxylic acid cycle (TCA cycle) itaconic acid. By optimizing PCN, could achieve titer 3 g/L, 5.3-fold higher than strain. Conclusions Our offers identify genes competitive relationship with pathways crucial growth organism. approach can potentially be applied other bacterial hosts substituting sensing module or gene.

Язык: Английский

Процитировано

0