Developing Riboswitch-Mediated Gene Regulatory Controls in Thermophilic Bacteria

Katherine Chou, Jonathan Lo, Ambarish Nag, Pin-Ching Maness, Joan Marcano

Research output: Contribution to journalArticlepeer-review

25 Scopus Citations

Abstract

Thermophilic bacteria are attractive hosts to produce bio-based chemicals. While various genetic manipulations have been employed in the metabolic engineering of thermophiles, a robust means to regulate gene expression in these bacteria (∼55 °C) is missing. Our bioinformatic search for various riboswitches in thermophilic bacteria revealed that major classes of riboswitches are present, suggesting riboswitches' regulatory roles in these bacteria. By building synthetic constructs incorporating natural and engineered purine riboswitch sequences originated from foreign species, we quantified respective riboswitches activities in repressing and up-regulating gene expression in Geobacillus thermoglucosidasius using a green fluorescence protein. The elicited regulatory response was ligand-concentration-dependent. We further demonstrated that riboswitch-mediated gene expression of adhE (responsible for ethanol production) in Clostridium thermocellum can modulate ethanol production, redirect metabolites, and control cell growth in the adhE knockout mutant. This work has made tunable gene expression feasible across different thermophiles for broad applications including biofuels production and gene-to-trait mapping.

Original languageAmerican English
Pages (from-to)633-640
Number of pages8
JournalACS Synthetic Biology
Volume8
Issue number4
DOIs
StatePublished - 19 Apr 2019

Bibliographical note

Publisher Copyright:
© 2019 American Chemical Society.

NREL Publication Number

  • NREL/JA-2700-73607

Keywords

  • Clostridium thermocellum
  • gene regulation
  • inducible and repressible systems
  • riboswitch
  • thermophiles

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