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Design, development and application of whole-cell based antibiotic-specific biosensor.

Yuriy Rebets1, Stefan Schmelz2, Oleksandr Gromyko3

  • 1Universität des Saarlandes, Pharmazeutische Biotechnologie, Bld. C2.3, 66123 Saarbrücken, Germany; Helmholtz-Institute for Pharmaceutical Research Saarland, UdS Campus, Bld. E8.1, 66123 Saarbrücken, Germany.

Metabolic Engineering
|April 3, 2018
PubMed
Summary

We developed novel whole-cell biosensors to improve antibiotic production in challenging actinobacteria. These engineered biosensors enhance the identification and optimization of natural product drug discovery.

Keywords:
ActinobacteriaAntibioticPolyketideSecondary metaboliteTetR repressor proteinWhole cell biosensor

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Area of Science:

  • Synthetic biology
  • Microbial biotechnology
  • Natural product drug discovery

Background:

  • Optimizing natural product synthesis in microorganisms is crucial for drug discovery.
  • Evaluating modified bacterial phenotypes, especially in slow-growing actinobacteria, is a significant bottleneck.
  • Actinobacteria are genetically challenging to manipulate, hindering antibiotic production enhancement.

Purpose of the Study:

  • To engineer antibiotic-specific whole-cell biosensors for identifying and optimizing antibiotic-producing strains.
  • To overcome limitations in dynamic and operating ranges of initial biosensor designs.
  • To provide recommendations for constructing and applying biosensors in actinobacteria.

Main Methods:

  • Generation of antibiotic-specific whole-cell biosensors using the TetR transcriptional repressor system.
  • Fine-tuning biosensor performance by modifying promoter, operator, and transcription factor ligand affinity.
  • Application of the biosensor to improve polyketide antibiotic pamamycin production.

Main Results:

  • Successfully generated and applied a TetR-derived whole-cell biosensor.
  • Improved the production of the polyketide antibiotic pamamycin using the biosensor.
  • Identified limitations in initial biosensor designs and implemented modifications to enhance performance.

Conclusions:

  • Developed a functional whole-cell biosensor for optimizing antibiotic production in actinobacteria.
  • Demonstrated the utility of biosensor fine-tuning for improved dynamic and operating ranges.
  • Provided key recommendations for the design and application of actinobacterial biosensors in synthetic biology.