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Microbial Biosensors01:17

Microbial Biosensors

Microbial biosensors are analytical devices that utilize living microbes to detect specific substances through measurable signals. These devices consist of two main components: biosensing organisms and signal-transducing elements. Biosensing organisms, such as Escherichia coli or Saccharomyces cerevisiae, are typically housed in multiwell plates connected to transducers, enabling rapid, real-time detection of target analytes.Signal Generation MechanismWhen a target analyte—such as...

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Biosensor development for single-cell detection of glucuronate.

Jennifer Kaczmarek Nash1, Kristala L J Prather1

  • 1Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139,USA.

Journal of Industrial Microbiology & Biotechnology
|June 16, 2023
PubMed
Summary

This study optimized a transcription-factor biosensor for screening myo-inositol oxygenase (MIOX) variants. Modifications improved biosensor performance, enabling the separation of closely related MIOX homologs.

Keywords:
BiosensorsDirected evolutionMetabolic engineeringMetabolite-responsive transcription factorSynthetic biology

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Enzyme Engineering

Background:

  • Biosensors offer high-throughput screening capabilities for genetic libraries.
  • Physiological limitations and mechanistic knowledge gaps can hinder biosensor application, similar to microbial systems.
  • Transcription-factor-based biosensors, like the ExuR system, are valuable tools in synthetic biology.

Purpose of the Study:

  • To characterize a previously developed transcription-factor (ExuR) based galacturonate biosensor for its cognate ligand, glucuronate.
  • To investigate the biosensor's performance with different myo-inositol oxygenase (MIOX) homologs.
  • To optimize biosensor circuit architecture and culture conditions for improved performance and application in MIOX variant screening.

Main Methods:

  • Characterization of an ExuR-based biosensor for glucuronate.
  • Testing biosensor response across various MIOX homologs.
  • Modification of biosensor circuit architecture and culture conditions.
  • Application of the optimized biosensor for separating closely related MIOX homologs.

Main Results:

  • The ExuR-based biosensor showed an ideal response to glucuronate under controlled conditions.
  • Biosensor performance deviated when applied to different MIOX homologs, indicating system variability.
  • Circuit architecture and culture condition modifications successfully reduced variation.
  • The optimized biosensor was effectively used to separate two closely related MIOX homologs.

Conclusions:

  • The ExuR-based biosensor can be adapted and optimized for screening MIOX variants.
  • Modifications to biosensor design and culture conditions are crucial for robust performance in complex biological applications.
  • This work demonstrates the utility of biosensors in enzyme engineering and the separation of homologous proteins.