Reversible thermal regulation for bifunctional dynamic control of gene expression in Escherichia coli

Nat Commun. 2021 Mar 3;12(1):1411. doi: 10.1038/s41467-021-21654-x.

Abstract

Genetically programmed circuits allowing bifunctional dynamic regulation of enzyme expression have far-reaching significances for various bio-manufactural purposes. However, building a bio-switch with a post log-phase response and reversibility during scale-up bioprocesses is still a challenge in metabolic engineering due to the lack of robustness. Here, we report a robust thermosensitive bio-switch that enables stringent bidirectional control of gene expression over time and levels in living cells. Based on the bio-switch, we obtain tree ring-like colonies with spatially distributed patterns and transformer cells shifting among spherical-, rod- and fiber-shapes of the engineered Escherichia coli. Moreover, fed-batch fermentations of recombinant E. coli are conducted to obtain ordered assembly of tailor-made biopolymers polyhydroxyalkanoates including diblock- and random-copolymer, composed of 3-hydroxybutyrate and 4-hydroxybutyrate with controllable monomer molar fraction. This study demonstrates the possibility of well-organized, chemosynthesis-like block polymerization on a molecular scale by reprogrammed microbes, exemplifying the versatility of thermo-response control for various practical uses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Hydroxybutyric Acid / metabolism
  • Escherichia coli / genetics*
  • Escherichia coli / growth & development
  • Escherichia coli / metabolism
  • Fermentation
  • Gene Expression Regulation, Bacterial*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Hydroxybutyrates / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Metabolic Engineering / methods*
  • Microorganisms, Genetically-Modified
  • Polyesters / metabolism
  • Polyhydroxyalkanoates / metabolism*
  • Red Fluorescent Protein
  • Temperature
  • Time-Lapse Imaging

Substances

  • Hydroxybutyrates
  • Luminescent Proteins
  • Polyesters
  • Polyhydroxyalkanoates
  • poly(3-hydroxybutyrate)-block-poly(4-hydroxybutyrate)
  • poly(3-hydroxypropionate-co-4-hydroxybutyrate)
  • Green Fluorescent Proteins
  • 3-Hydroxybutyric Acid