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Engineering protein production by rationally choosing a carbon and nitrogen source using E. coli BL21 acetate metabolism knockout strains.

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dc.contributor.authorLozano Terol, Gema
dc.contributor.authorCánovas Díaz, Manuel
dc.contributor.authorDiego Puente, Teresa de
dc.contributor.authorGallego Jara, Julia
dc.contributor.authorSola Martínez, Rosa Alba
dc.contributor.departmentBioquímica y Biología Molecular B e Inmunología
dc.date.accessioned2023-12-12T23:11:56Z
dc.date.available2023-12-12T23:11:56Z
dc.date.issued2019-09-04
dc.description©2019. The authors. This document is made available under the CC-BY 4.0 license http://creativecommons.org/licenses/by /4.0/ This document is the published version of a Published Work that appeared in final form in Microbial Cell Factories. To access the final edited and published work see https://doi.org/10.1186/s12934-019-1202-1
dc.description.abstractBackground: Escherichia 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 overfow. 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 fnd the most favourable conditions for carrying out recombinant protein production in E. coli BL21 using two diferent approaches, namely, manipulation of the culture media composition and the deletion of genes involved in acetate metabolism and Nε-lysine acetylation. Results: We evaluated protein overexpression in E. coli BL21 wt and fve 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 efcient use of resources, since is a sub-product of biodiesel synthesis. Furthermore, the deletion of the ackA gene results in a fvefold increase in protein production with respect to the wt strain and a reduction in acetate accumulation. Conclusion: The results showed that the use of diverse carbon and nitrogen sources and acetate metabolism knockout strains can redirect E. coli carbon fuxes to diferent pathways and afect the fnal yield of the recombinant protein bioprocess. Thereby, we obtained a fvefold increase in protein production and an efcient use of the resources employing the most suitable strain and culture conditions.
dc.formatapplication/pdfes_ES
dc.format.extent19
dc.identifier.citationMicrobial Cell Factories (2019)18:151
dc.identifier.doihttps://doi.org/10.1186/s12934-019-1202-1
dc.identifier.issn1475-2859
dc.identifier.urihttp://hdl.handle.net/10201/136583
dc.languageenges_ES
dc.publisherSpringer Nature
dc.relationThis work was supported by grants from the Ministry of Science, Innovation and Universities (MCIU), the State Research Agency (AEI) and the European Regional Development Fund (FEDER), RTI2018-094393-B-C21-MCIU/AEI/ FEDER, UE, and the Seneca Foundation CARM, 20786/PI/18
dc.relation.isreferencedbyED_IDENTRADA=1274
dc.relation.publisherversionhttps://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-019-1202-1
dc.rightsinfo:eu-repo/semantics/openAccess*
dc.rightsAtribución 4.0 Internacional*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectRecombinant proteins
dc.subjectAcetate overfow
dc.subjectEscherichia coli
dc.subjectLysine acetylation
dc.subjectGFP
dc.subjectMedium composition
dc.titleEngineering protein production by rationally choosing a carbon and nitrogen source using E. coli BL21 acetate metabolism knockout strains.es_ES
dc.typeinfo:eu-repo/semantics/articlees_ES
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relation.isAuthorOfPublication.latestForDiscovery57bf3537-1124-49ab-8774-bd0c3bdf1c28
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