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

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  • معلومة اضافية
    • المصدر:
      Publisher: BioMed Central Country of Publication: England NLM ID: 101139812 Publication Model: Electronic Cited Medium: Internet ISSN: 1475-2859 (Electronic) Linking ISSN: 14752859 NLM ISO Abbreviation: Microb Cell Fact Subsets: MEDLINE
    • بيانات النشر:
      Original Publication: London : BioMed Central, [2002-
    • الموضوع:
      Metabolic Engineering*; Alcohols/*metabolism ; Escherichia coli/*metabolism ; Esters/*metabolism ; Fatty Acids/*metabolism; Alcohols/chemistry ; Bioreactors ; Biosynthetic Pathways ; Coculture Techniques ; Escherichia coli/genetics ; Escherichia coli/growth & development ; Esters/chemistry ; Fatty Acids/chemistry ; Fermentation ; Glucose/metabolism ; Methylation ; Propionates/metabolism
    • نبذة مختصرة :
      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.
      (© 2022. The Author(s).)
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    • Grant Information:
      PICT-2017-2243 Agencia Nacional de Promoción Científica y Tecnológica; PICT-2017-2184 Agencia Nacional de Promoción Científica y Tecnológica
    • Contributed Indexing:
      Keywords: E. coli coculture; Fed-batch culture; Methyl-branched alcohols; Multi-methyl-branched esters; Multi-methyl-branched fatty acids; Response surface methodology
    • الرقم المعرف:
      0 (Alcohols)
      0 (Esters)
      0 (Fatty Acids)
      0 (Propionates)
      IY9XDZ35W2 (Glucose)
    • الموضوع:
      Date Created: 20220116 Date Completed: 20220121 Latest Revision: 20231105
    • الموضوع:
      20250114
    • الرقم المعرف:
      PMC8760833
    • الرقم المعرف:
      10.1186/s12934-022-01737-0
    • الرقم المعرف:
      35033081