ArticleMicrobial cell factories2019
Engineering protein production by rationally choosing a carbon and nitrogen source using E. coli BL21 acetate metabolism knockout strains.
Article in Microbial cell factories, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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29 citing papers in PubMed, 69 citations in OpenAlex.
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- Overflow metabolism in bacterial, yeast, and mammalian cells: different names, same game.Molecular systems biology · 2025Review
- Algebraic differentiation for fast sensitivity analysis of optimal flux modes in metabolic models.Bioinformatics (Oxford, England) · 2025Article
- Induction conditions that promote the effect of glycerol on recombinant protein production inBiotechnology reports (Amsterdam, Netherlands) · 2025Article
- Cra-controlled antisense RNA-downregulation of isocitrate dehydrogenase in Escherichia coli.Archives of microbiology · 2025Article
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- Modeling and optimization of culture media for recombinantFrontiers in bioengineering and biotechnology · 2024Article
- Optimization of hydrogen production in Enterobacter aerogenes by Complex I peripheral fragments destruction and maeA overexpression.Microbial cell factories · 2023Article
- Development of high-copy number plasmids in Pseudoalteromonas haloplanktis TAC125.Applied microbiology and biotechnology · 2023Article
- Regulation of the pyrimidine biosynthetic pathway by lysine acetylation of E. coli OPRTase.The FEBS journal · 2023Article
- High-level recombinant protein production with Corynebacterium glutamicum using acetate as carbon source.Microbial biotechnology · 2022Article
- Strategies for efficient production of recombinant proteins in Escherichia coli: alleviating the host burden and enhancing protein activity.Microbial cell factories · 2022Review
- An Esterase with Increased Acetone Tolerance from Bacillus subtilis E9 over Expressed in E. coli BL21 Using pTac Bs-est Vector.Molecular biotechnology · 2022Article
- Heme A Synthase Deficiency Affects the Ability ofInternational journal of molecular sciences · 2022Article
- Advances and prospects in metabolic engineering of Escherichia coli for L-tryptophan production.World journal of microbiology & biotechnology · 2022Review
- Model-based dynamic engineering ofBiotechnology notes (Amsterdam, Netherlands) · 2022Article
- Optimizing the Expression and Solubilization of an E. coli-Produced Leukemia Inhibitory Factor for Anti-LIF Antibody Production and Use Thereof for Contraception in Mice.Molecular biotechnology · 2021Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
backgroundEscherichia coli (E. coli) is a bacteria that is widely employed in many industries for the production of high interest bio-products such as recombinant proteins. Nevertheless, the use of E. coli for recombinant protein production may entail some disadvantages such as acetate overflow. Acetate is accumulated under some culture conditions, involves a decrease in biomass and recombinant protein production, and its metabolism is related to protein lysine acetylation. Thereby, the carbon and nitrogen sources employed are relevant factors in cell host metabolism, and the study of the central metabolism of E. coli and its regulation is essential for optimizing the production of biomass and recombinant proteins. In this study, our aim was to find the most favourable conditions for carrying out recombinant protein production in E. coli BL21 using two different approaches, namely, manipulation of the culture media composition and the deletion of genes involved in acetate metabolism and Nε-lysine acetylation.
resultsWe evaluated protein overexpression in E. coli BL21 wt and five mutant strains involved in acetate metabolism (Δacs, ΔackA and Δpta) and lysine acetylation (ΔpatZ and ΔcobB) grown in minimal medium M9 (inorganic ammonium nitrogen source) and in complex TB7 medium (peptide-based nitrogen source) supplemented with glucose (PTS carbon source) or glycerol (non-PTS carbon source). We observed a dependence of recombinant protein production on acetate metabolism and the carbon and nitrogen source employed. The use of complex medium supplemented with glycerol as a carbon source entails an increase in protein production and an efficient use of resources, since is a sub-product of biodiesel synthesis. Furthermore, the deletion of the ackA gene results in a fivefold increase in protein production with respect to the wt strain and a reduction in acetate accumulation.
conclusionThe results showed that the use of diverse carbon and nitrogen sources and acetate metabolism knockout strains can redirect E. coli carbon fluxes to different pathways and affect the final yield of the recombinant protein bioprocess. Thereby, we obtained a fivefold increase in protein production and an efficient use of the resources employing the most suitable strain and culture conditions.
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