Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids
Background: 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 th...
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| Autores principales: | , , , |
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| Formato: | article artículo publishedVersion |
| Lenguaje: | Inglés |
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BioMed Central Ltd
2022
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| Acceso en línea: | http://hdl.handle.net/2133/23376 http://hdl.handle.net/2133/23376 |
| Aporte de: |
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I15-R121-2133-23376 |
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dspace |
| institution |
Universidad Nacional de Rosario |
| institution_str |
I-15 |
| repository_str |
R-121 |
| collection |
Repositorio Hipermedial de la Universidad Nacional de Rosario (UNR) |
| language |
Inglés |
| orig_language_str_mv |
eng |
| topic |
E. coli coculture Multi-methyl-branched fatty acids Methyl-branched alcohols Multi-methyl-branched esters Fed-batch culture Response surface methodology |
| spellingShingle |
E. coli coculture Multi-methyl-branched fatty acids Methyl-branched alcohols Multi-methyl-branched esters Fed-batch culture Response surface methodology Bracalente, Fernando Sabatini, Martín Arabolaza, Ana Lorena Gramajo, Hugo Cesar Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids |
| topic_facet |
E. coli coculture Multi-methyl-branched fatty acids Methyl-branched alcohols Multi-methyl-branched esters Fed-batch culture Response surface methodology |
| description |
Background: 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. |
| format |
article artículo publishedVersion |
| author |
Bracalente, Fernando Sabatini, Martín Arabolaza, Ana Lorena Gramajo, Hugo Cesar |
| author_facet |
Bracalente, Fernando Sabatini, Martín Arabolaza, Ana Lorena Gramajo, Hugo Cesar |
| author_sort |
Bracalente, Fernando |
| title |
Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids |
| title_short |
Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids |
| title_full |
Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids |
| title_fullStr |
Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids |
| title_full_unstemmed |
Escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids |
| title_sort |
escherichia coli coculture for de novo production of esters derived of methyl-branched alcohols and multi-methyl branched fatty acids |
| publisher |
BioMed Central Ltd |
| publishDate |
2022 |
| url |
http://hdl.handle.net/2133/23376 http://hdl.handle.net/2133/23376 |
| work_keys_str_mv |
AT bracalentefernando escherichiacolicoculturefordenovoproductionofestersderivedofmethylbranchedalcoholsandmultimethylbranchedfattyacids AT sabatinimartin escherichiacolicoculturefordenovoproductionofestersderivedofmethylbranchedalcoholsandmultimethylbranchedfattyacids AT arabolazaanalorena escherichiacolicoculturefordenovoproductionofestersderivedofmethylbranchedalcoholsandmultimethylbranchedfattyacids AT gramajohugocesar escherichiacolicoculturefordenovoproductionofestersderivedofmethylbranchedalcoholsandmultimethylbranchedfattyacids |
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Repositorios |
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