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Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
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Tuning Extracellular Electron Transfer by Shewanella oneidensis Using Transcriptional Logic Gates.

Christopher M Dundas1, David J F Walker2, Benjamin K Keitz1

  • 1McKetta Department of Chemical Engineering, University of Texas at Austin, Austin, Texas 78712, United States.

ACS Synthetic Biology
|August 14, 2020
PubMed
Summary

Researchers engineered the bacterium Shewanella oneidensis for better control over extracellular electron transfer (EET). This work enables predictable programming of EET flux using transcriptional logic gates for redox-driven applications.

Keywords:
Shewanella oneidensisextracellular electron transferlogic gate

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Area of Science:

  • Microbiology
  • Synthetic Biology
  • Biochemistry

Background:

  • Extracellular electron transfer (EET) pathways in bacteria like Shewanella oneidensis are crucial for interfacing cellular metabolism with redox applications.
  • Controlling EET flux in S. oneidensis is challenging due to limited understanding of pathway proteins and their impact on engineering parameters.

Purpose of the Study:

  • To systematically investigate how altering transcription and translation of key EET pathway genes (CymA/MtrCAB) affects Fe(III) reduction kinetics in S. oneidensis.
  • To develop engineered control over EET flux using transcriptional logic gates.

Main Methods:

  • Systematic alteration of transcription and translation of single genes encoding CymA, MtrA, and MtrC in S. oneidensis.
  • Utilized plasmid-based inducible circuits in S. oneidensis knockout strains to modulate gene expression.
  • Developed Buffer and NOT gate architectures responsive to isopropyl β-D-1-thiogalactopyranoside (IPTG) to control EET flux.

Main Results:

  • Identified specific construct/strain pairings that maximize the dynamic range of Fe(III) reduction rate by optimizing cymA, mtrA, and mtrC expression.
  • Demonstrated predictable ON/OFF control of EET flux using engineered Buffer and NOT gates.
  • Response functions of engineered logic gates showed comparability to conventional synthetic biology circuits with fluorescent reporters.

Conclusions:

  • Provides insights into programming EET activity using transcriptional logic gates.
  • Suggests adaptability of existing transcriptional circuitry for predictable control of EET flux in S. oneidensis.
  • Advances the engineering of microbial electrochemical systems and related redox-driven applications.