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Generation of a platform strain for ionic liquid tolerance using adaptive laboratory evolution.

Elsayed T Mohamed1, Shizeng Wang2,3,4, Rebecca M Lennen1

  • 1Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Microbial Cell Factories
|November 18, 2017
PubMed
Summary
This summary is machine-generated.

Engineered *Escherichia coli* strains exhibit enhanced tolerance to toxic ionic liquids (ILs) after adaptive laboratory evolution. These improved microbial platforms are crucial for sustainable bioproduction from biomass-derived sugars.

Keywords:
Adaptive laboratory evolutionEscherichia coliIonic liquidsRenewable feedstocks

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Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Microbial Engineering

Background:

  • Petroleum-derived chemicals face sustainability challenges, necessitating alternatives from renewable biomass.
  • Ionic liquids (ILs) are effective for biomass deconstruction but exhibit toxicity to microbial fermentation.
  • Residual IL toxicity hinders the bioprocessing of sugars derived from IL-treated biomass.

Purpose of the Study:

  • To develop *Escherichia coli* strains tolerant to residual ionic liquids (ILs) for improved bioproduction.
  • To overcome the toxicity limitations of ILs in biomass conversion processes.

Main Methods:

  • A tolerance adaptive laboratory evolution (TALE) approach was employed using *Escherichia coli* strains DH1 and K-12 MG1655.
  • Strains were evolved over approximately 40 days in the presence of specific ILs: 1-ethyl-3-methylimidazolium acetate ([C2C1Im][OAc]) and 1-butyl-3-methylimidazolium chloride ([C4C1Im]Cl).
  • Whole genome sequencing was performed on evolved clonal isolates to identify genetic modifications.

Main Results:

  • Tolerant *E. coli* clones exhibited mutations in multidrug efflux pumps (mdtJI) and uncharacterized transporters (yhdP).
  • Enriched mutations were also observed in transcriptional regulation and nucleotide biosynthesis pathways.
  • The best-performing evolved strains demonstrated robust growth at 8.5% (w/v) and detectable growth up to 11.9% (w/v) [C2C1Im][OAc], showing cross-tolerance.

Conclusions:

  • The generated *E. coli* strains are superior platform strains for bioproduction using IL-treated renewable feedstocks.
  • The TALE method effectively addresses IL substrate toxicity, showing promise for broader tolerance engineering applications.