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

et al.

Journal of Biological Engineering, Journal Year: 2024, Volume and Issue: 18(1)

Published: Dec. 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.

Language: Английский

Recent advances in the Design-Build-Test-Learn (DBTL) cycle for systems metabolic engineering of Corynebacterium glutamicum DOI

Subeen Jeon,

Yu Jung Sohn,

Haeyoung Lee

et al.

The Journal of Microbiology, Journal Year: 2025, Volume and Issue: 63(3), P. e2501021 - e2501021

Published: March 28, 2025

Language: Английский

Citations

1
Enhancement Strategy for Protocatechuic Acid Production Using Corynebacterium glutamicum with Focus on Continuous Fermentation Scale-Up and Cytotoxicity Management DOI Open Access

J. Chung,

Woo-Shik Shin, Chulhwan Park

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(1), P. 396 - 396

Published: Jan. 5, 2025

Protocatechuate acid (PCA) is a phenolic naturally synthesized by various organisms. Protocatechuic plants for physiological, metabolic functions, and self-defense, but extraction from less efficient compared to the microbial culture process. The synthesis of protocatechuic sustainable and, due its high yield, can save energy consumption when producing same amount. To enhance PCA production using Corynebacterium glutamicum, statistical optimization medium was performed full factorial design, steepest ascent method, response surface method. optimized enabled over 5 g/L in 72 h batch culture. However, cytotoxicity affected strain growth rate, with an inhibitory concentration approximately fermentation broth. Finally, continuous operated 150 steady-state mode, maintaining below g/L. method established this study successfully increased levels, findings presented herein are anticipated contribute industrialization C. glutamicum.

Language: Английский

Citations

0
Advances in engineering and applications of microbial glutamate decarboxylases for gamma-aminobutyric acid production DOI

Yingjie Lu,

Feng Zhang, Leli Wang

et al.

Enzyme and Microbial Technology, Journal Year: 2025, Volume and Issue: 188, P. 110652 - 110652

Published: April 6, 2025

Language: Английский

Citations

0
Review of the Proteomics and Metabolic Properties of Corynebacterium glutamicum DOI Creative Commons

J M Park,

Sooa Lim

Microorganisms, Journal Year: 2024, Volume and Issue: 12(8), P. 1681 - 1681

Published: Aug. 15, 2024

Corynebacterium glutamicum (C. glutamicum) has become industrially important in producing glutamic acid and lysine since its discovery been the subject of proteomics central carbon metabolism studies. The proteome changes depending on environmental conditions, nutrient availability, stressors. Post-translational modification (PTMs), such as phosphorylation, methylation, glycosylation, alter function activity proteins, allowing them to respond quickly changes. Proteomics techniques, mass spectrometry two-dimensional gel electrophoresis, have enabled study proteomes, identification quantification expression levels. Understanding proteomes microorganisms provides insight into their physiology, ecology, biotechnological applications, biofuels, pharmaceuticals, industrial enzyme production. Several attempts made create efficient production strains increase productivity several research fields, genomics proteomics. In addition amino acids, C. is used produce vitamins, nucleotides, organic alcohols, expanding applications. Considerable information accumulated, but recent focused metabolism. development genetic engineering technologies, CRISPR-Cas9, improved efficiency by precise manipulation metabolic pathways glutamicum. addition, methods for designing new developing customized using synthetic biology technology are gradually expanding. This review expected enhance understanding potential help researchers identify topics design

Language: Английский

Citations

1
Efficient Synthesis of γ-Aminobutyric Acid from Monosodium Glutamate Using an Engineered Glutamate Decarboxylase Active at a Neutral pH DOI Open Access
Miao Li,

Yingjia Zheng,

Ran Cheng

et al.

Catalysts, Journal Year: 2024, Volume and Issue: 14(12), P. 905 - 905

Published: Dec. 9, 2024

The efficient production of γ-aminobutyric acid (GABA) at a neutral pH remains challenge due to the sensitivity glutamate decarboxylase (GAD) enzymes. Our study addressed this limitation by identifying and engineering GAD enzymes with high activity under conditions. Through gene mining, we discovered wild-type from Enterococcus faecalis (EfGAD) 7.0 and, using zero-shot (ZS) predictor-guided mutagenesis C-terminal truncation, developed an EfGAD variant significantly enhanced catalytic efficiency. This demonstrated 1.3-fold increase in GABA (~300 g/L) monosodium (MSG) compared 5 L bioreactor experiments. ability operate without need for acidic conditions reduces costs facilitates scalability. findings underscore potential integrating machine learning tools enzyme optimization provide sustainable approach biosynthesis MSG as substrate.

Language: Английский

Citations

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

et al.

Journal of Biological Engineering, Journal Year: 2024, Volume and Issue: 18(1)

Published: Dec. 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.

Language: Английский

Citations

0