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Developing cell factories for sustainable chemical production requires efficient strain screening. Researchers engineered a transcriptional regulator biosensor for ultra-high-throughput screening of L-histidine producers using fluorescence-activated cell sorting.

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Area of Science:

  • Metabolic Engineering
  • Synthetic Biology
  • Biocatalysis

Background:

  • Cell factories offer a sustainable route to chemical production, but screening microbial strains is inefficient.
  • Transcriptional biosensors coupled with fluorescence-activated cell sorting (FACS) enable high-throughput screening.
  • Broad ligand specificity of transcriptional regulators (TRs) limits the development of precise biosensors.

Purpose of the Study:

  • To engineer a specific biosensor for L-histidine production by modifying the LysG transcriptional regulator.
  • To overcome the limitation of broad ligand specificity in TRs for enhanced biosensor applications.

Main Methods:

  • Determined the structure of the LysG transcriptional regulator from Corynebacterium glutamicum.
  • Employed a semi-rational engineering approach combined with FACS-based screening and counterscreening.
  • Generated a LysG-based biosensor with reduced sensitivity to L-lysine.

Main Results:

  • Successfully engineered a LysG-based biosensor that is insensitive to L-lysine.
  • Demonstrated the utility of the engineered biosensor for isolating L-histidine-producing strains via FACS.
  • Validated that TR engineering can narrow ligand specificity, expanding metabolite sensor applications.

Conclusions:

  • TR engineering is a viable strategy to create specific biosensors for ultra-high-throughput screening.
  • The developed L-lysine-insensitive LysG biosensor expands the toolkit for metabolic engineering and synthetic biology.
  • This work facilitates the development of more efficient cell factories for sustainable chemical production.