Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids

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dc.citation.titleMicrobial Cell Factorieses
dc.citation.volume21
dc.creatorBracalente, Fernando
dc.creatorSabatini, Martín
dc.creatorArabolaza, Ana Lorena
dc.creatorGramajo, Hugo Cesar
dc.date.accessioned2022-04-08T16:12:19Z
dc.date.available2022-04-08T16:12:19Z
dc.date.issued2022-01-15
dc.descriptionBackground: A broad diversity of natural and non-natural esters have now been made in bacteria, and in other microorganisms, as a result of original metabolic engineering approaches. However, the fact that the properties of these molecules, and therefore their applications, are largely defined by the structural features of the fatty acid and alcohol moieties, has driven a persistent interest in generating novel structures of these chemicals. Results: In this research, we engineered Escherichia coli to synthesize de novo esters composed of multi-methyl-branched-chain fatty acids and short branched-chain alcohols (BCA), from glucose and propionate. A coculture engineering strategy was developed to avoid metabolic burden generated by the reconstitution of long heterologous biosynthetic pathways. The cocultures were composed of two independently optimized E. coli strains, one dedicated to efficiently achieve the biosynthesis and release of the BCA, and the other to synthesize the multi methyl-branched fatty acid and the corresponding multi-methyl-branched esters (MBE) as the final products. Response surface methodology, a cost-efficient multivariate statistical technique, was used to empirical model the BCA-derived MBE production landscape of the coculture and to optimize its productivity. Compared with the monoculture strategy, the utilization of the designed coculture improved the BCA-derived MBE production in 45%. Finally, the coculture was scaled up in a high-cell density fed-batch fermentation in a 2 L bioreactor by fine-tuning the inoculation ratio between the two engineered E. coli strains. Conclusion: Previous work revealed that esters containing multiple methyl branches in their molecule present favorable physicochemical properties which are superior to those of linear esters. Here, we have successfully engineered an E. coli strain to broaden the diversity of these molecules by incorporating methyl branches also in the alcohol moiety. The limited production of these esters by a monoculture was considerable improved by a design of a coculture system and its optimization using response surface methodology. The possibility to scale-up this process was confirmed in high-cell density fed-batch fermentations.es
dc.description.filFil: Bracalente, Fermando. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas Y Farmacéuticas. Microbiology Division. Instituto de Biología Molecular Y Celular de Rosario (IBR-CONICET); Argentina.es
dc.description.filFil: Sabatini, Martín. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas Y Farmacéuticas. Microbiology Division. Instituto de Biología Molecular Y Celular de Rosario (IBR-CONICET); Argentina.es
dc.description.filFil: Arabolaza, Ana Lorena. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas Y Farmacéuticas. Microbiology Division. Instituto de Biología Molecular Y Celular de Rosario (IBR-CONICET); Argentina.es
dc.description.filFil: Gramajo, Hugo Cesar. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas Y Farmacéuticas. Microbiology Division. Instituto de Biología Molecular Y Celular de Rosario (IBR-CONICET); Argentina.es
dc.description.sponsorshipAgencia Nacional de Promoción de la Investigación. Desarrollo Tecnológico y la Innovación (ANPCyT): PICT-2017–2243, PICT-2017–2184
dc.description.sponsorshipAgencia Nacional de Promoción de la Investigación. Desarrollo Tecnológico y la Innovación (ANPCyT): PICT-2017–2243, PICT-2017–2184
dc.formatapplication/pdf
dc.format.extent1-17es
dc.identifier.issn1475-2859es
dc.identifier.urihttp://hdl.handle.net/2133/23376
dc.language.isoenges
dc.publisherBioMed Central Ltdes
dc.relation.publisherversionhttps://doi.org/10.1186/s12934-022-01737-0
dc.relation.publisherversionhttps://microbialcellfactories.biomedcentral.com/track/pdf/10.1186/s12934-022-01737-0.pdf
dc.rightsopenAccesses
dc.rights.holderBracalente, Fernandoes
dc.rights.holderSabatini, Martínes
dc.rights.holderArabolaza, Ana Lorenaes
dc.rights.holderGramajo, Hugo Cesares
dc.rights.textAttribution 4.0 International (CC BY 4.0)es
dc.subjectE. coli coculturees
dc.subjectMulti-methyl-branched fatty acidses
dc.subjectMethyl-branched alcoholses
dc.subjectMulti-methyl-branched esterses
dc.subjectFed-batch culturees
dc.subjectResponse surface methodologyes
dc.titleEscherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acidses
dc.typepublishedVersion
dc.typearticle
dc.typeartículo
dc.type.collectionarticulo
dc.type.versionpublishedVersion

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