Precise precursor rebalancing for isoprenoids production by fine control of gapA expression in Escherichia coli

Jung, Ju‐Young; Lim, Jae Hyung; Kim, Se Yeon; Im, Dae Kyun; Seok, Joo Yeon; Lee, Seung-Jae; Oh, Min Kyu; Jung, Gyoo Yeol

doi:10.1016/j.ymben.2016.10.003

Cited by 55 publications

(52 citation statements)

References 40 publications

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“…It has been documented that ptsG mutants showed higher production of lycopene compared to the parental strain, however, the mechanism was not suggested in a prior study [68]. Another study demonstrated that activation of ppsA and repression of gapA were attempted to balance intracellular G3P and pyruvate, precursors of MEP pathway [69]. According to our transcriptome analysis, transcription of gapA was downregulated and that of ppsA was upregulated in the sugar transporter mutants.…”

Section: Application Of the Mutants For Metabolite Production (Egfp mentioning

confidence: 54%

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Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Jung

Lim

et al. 2019

Microb Cell Fact

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Background: Most microorganisms have evolved to maximize growth rate, with rapid consumption of carbon sources from the surroundings. However, fast growing phenotypes usually feature secretion of organic compounds. For example, E. coli mainly produced acetate in fast growing condition such as glucose rich and aerobic condition, which is troublesome for metabolic engineering because acetate causes acidification of surroundings, growth inhibition and decline of production yield. The overflow metabolism can be alleviated by reducing glucose uptake rate. Results: As glucose transporters or their subunits were knocked out in E. coli, the growth and glucose uptake rates decreased and biomass yield was improved. Alteration of intracellular metabolism caused by the mutations was investigated with transcriptome analysis and 13 C metabolic flux analysis (13 C MFA). Various transcriptional and metabolic perturbations were identified in the sugar transporter mutants. Transcription of genes related to glycolysis, chemotaxis, and flagella synthesis was downregulated, and that of gluconeogenesis, Krebs cycle, alternative transporters, quorum sensing, and stress induced proteins was upregulated in the sugar transporter mutants. The specific production yields of value-added compounds (enhanced green fluorescent protein, γ-aminobutyrate, lycopene) were improved significantly in the sugar transporter mutants. Conclusions: The elimination of sugar transporter resulted in alteration of global gene expression and redirection of carbon flux distribution, which was purposed to increase energy yield and recycle carbon sources. When the pathways for several valuable compounds were introduced to mutant strains, specific yield of them were highly improved. These results showed that controlling the sugar uptake rate is a good strategy for ameliorating metabolite production.

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Section: Application Of the Mutants For Metabolite Production (Egfp mentioning

confidence: 54%

“…Jung et al [69] supernatant was removed after centrifugation (SMART R17; Hanil, Gimpo, Korea). About 1 mL of extraction solvent (mixture of equal volumes of methanol and acetone) was added to the pellet and mixed well.…”

Section: Methodsmentioning

confidence: 99%

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Jung

Lim

et al. 2019

Microb Cell Fact

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“…One OD 600 unit corresponds to 0.25 g dry cell weight (DCW)/L [34]. Instead, glucose consumption rate, determined as the analytic data from HPLC during the initial exponential phase, was represented for the production systems as the components in the TB medium also contributed to biomass yield.…”

Section: Methodsmentioning

confidence: 99%

A simple method to control glycolytic flux for the design of an optimal cell factory

Lim

Jung

2017

Biotechnol Biofuels

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BackgroundA microbial cell factory with high yield and productivity are prerequisites for an economically feasible bio-based chemical industry. However, cell factories that show a kinetic imbalance between glycolysis and product formation pathways are not optimal. Glycolysis activity is highly robust for survival in nature, but is not optimized for chemical production.ResultsHere, we propose a novel approach to balance glycolytic activity with the product formation capacity by precisely controlling expression level of ptsG (encoded glucose transporter) through UTR engineering. For various heterologous pathways with different maximum production rates, e.g., n-butanol, butyrate, and 2,3-butanediol, glycolytic fluxes could be successfully modulated to maximize yield and productivity, while minimizing by-product formation in Escherichia coli.ConclusionsThese results support the application of this simple method to explore the maximum yield and productivity when designing optimal cell factories for value-added products in the fields of metabolic engineering and synthetic biology.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0847-4) contains supplementary material, which is available to authorized users.

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“…Carotenoids are natural pigments present in plants and microorganisms [1]. Some carotenoids are known to function as membrane protective anti-oxidants [2,3], which has led to a high demand for carotenoids for medical and pharmaceutical applications [4,5]. Lycopene is one of the most highly valuable carotenoids owing to its potent antioxidant [6] and disease prevention properties [7,8].…”

Section: Introductionmentioning

confidence: 99%

“…Similarly, the catalytic activity of Dxs is known to be subjected to a negative feedback regulation by IPP and DMAPP [26], and expression of ispA (encoding farnesyl diphosphate synthase) could improve lycopene production by more than 2-fold [27]. On the other hand, there have been several attempts to increase lycopene production by balancing the expression levels of precursors, pyruvate, and G-3-P. Farmer and Liao were able to significantly improve lycopene production by a dynamic expression control of pps gene (encoding phosphoenolpyruvate synthase) [18]; similarly, Jung et al successfully demonstrated increased lycopene production through a precise expression control of gapA (encoding glyceraldehyde-3-phosphate dehydrogenase A) [4].…”

Section: Introductionmentioning

confidence: 99%

Enhanced Lycopene Production in Escherichia coli by Expression of Two MEP Pathway Enzymes from Vibrio sp. Dhg

et al. 2019

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Microbial production is a promising method that can overcome major limitations in conventional methods of lycopene production, such as low yields and variations in product quality. Significant efforts have been made to improve lycopene production by engineering either the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway or mevalonate (MVA) pathway in microorganisms. To further improve lycopene production, it is critical to utilize metabolic enzymes with high specific activities. Two enzymes, 1-deoxy-d-xylulose-5-phosphate synthase (Dxs) and farnesyl diphosphate synthase (IspA), are required in lycopene production using MEP pathway. Here, we evaluated the activities of Dxs and IspA of Vibrio sp. dhg, a newly isolated and fast-growing microorganism. Considering that the MEP pathway is closely related to the cell membrane and electron transport chain, the activities of the two enzymes of Vibrio sp. dhg were expected to be higher than the enzymes of Escherichia coli. We found that Dxs and IspA in Vibrio sp. dhg exhibited 1.08-fold and 1.38-fold higher catalytic efficiencies, respectively. Consequently, the heterologous overexpression improved the specific lycopene production by 1.88-fold. Our findings could be widely utilized to enhance production of lycopene and other carotenoids.

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Precise precursor rebalancing for isoprenoids production by fine control of gapA expression in Escherichia coli

Cited by 55 publications

References 40 publications

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

Metabolic perturbations in mutants of glucose transporters and their applications in metabolite production in Escherichia coli

A simple method to control glycolytic flux for the design of an optimal cell factory

Enhanced Lycopene Production in Escherichia coli by Expression of Two MEP Pathway Enzymes from Vibrio sp. Dhg

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