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Evolving a Generalist Biosensor for Bicyclic Monoterpenes.

Simon d'Oelsnitz1, Vylan Nguyen2, Hal S Alper3

  • 1Department of Molecular Biosciences, University of Texas at Austin, Austin, Texas 78712, United States.

ACS Synthetic Biology
|January 5, 2022
PubMed
Summary
This summary is machine-generated.

Scientists engineered versatile biosensors to detect specific monoterpenes, valuable compounds in flavors and fragrances. These tools enable precise chemical measurement and regulation within cells, advancing biotechnological applications.

Keywords:
biosensorsdirected evolutionmetabolic engineeringprotein engineeringterpenes

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Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Chemical Engineering

Background:

  • Monoterpenes are commercially valuable but challenging to measure in cellular environments.
  • Prokaryotic transcription factors offer potential as precise chemical measurement and regulation tools.

Purpose of the Study:

  • To develop genetically encoded, generalist monoterpene biosensors.
  • To expand the effector specificity of the TetR-family regulator CamR for monoterpene detection.

Main Methods:

  • Directed evolution was employed to modify the CamR regulator.
  • A novel screening method, Seamless Enrichment of Ligand-Inducible Sensors (SELIS), was utilized.
  • Promoter optimization was performed to enhance reporter gene expression.

Main Results:

  • Evolved CamR biosensors successfully recognized four distinct monoterpenes: borneol, fenchol, eucalyptol, and camphene.
  • Common mutations arose across different evolutionary paths, indicating CamR's platform utility.
  • Systematic promoter optimization achieved a 150-fold increase in signal-to-noise ratio.

Conclusions:

  • The developed CamR-based biosensors are effective tools for detecting specific monoterpenes.
  • These biosensors can be foundational for high-throughput screening and dynamic regulation of monoterpene production.
  • The engineered biosensors demonstrate a significant advancement in cellular chemical sensing technology.