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Rapid Optimization of a Light-Inducible System to Control Mammalian Gene Expression
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Genetic-Metabolic Coupling for Targeted Metabolic Engineering.

Stefano Cardinale1, Felipe Gonzalo Tueros1, Morten Otto Alexander Sommer1

  • 1NNF-CFB, Technical University of Denmark, 2800 Kongens Lyngby, Denmark.

Cell Reports
|August 3, 2017
PubMed
Summary

We developed a novel biosensor to link cell fitness with thiamine production in E. coli. This method identifies genetic targets affecting microbial chemical production and cell health.

Keywords:
Fe-S clustersRNA biosensorsoxidative stress responsethiamine biosynthesistransposon mutagenesis

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Area of Science:

  • Microbial biotechnology
  • Synthetic biology
  • Metabolic engineering

Background:

  • Microbial chemical production relies on enzymes that can impact cell fitness and yield.
  • Current optimization methods often overlook metabolic perturbations, limiting real-world applicability.

Purpose of the Study:

  • To develop a method linking cell fitness to thiamine diphosphate production in Escherichia coli.
  • To identify native gene networks influencing both cell fitness and thiamine production.

Main Methods:

  • Coupling cell fitness to thiamine diphosphate production using a synthetic RNA biosensor.
  • Interrogating a library of transposon mutants to elucidate the native gene network.
  • Identifying effectors of the OxyR-Fur stress response.

Main Results:

  • Identified effectors of the OxyR-Fur stress response that limit thiamine biosynthesis.
  • Demonstrated alternative regulation of iron storage and Fe-S cluster inclusion in enzymes.
  • Revealed a direct link between stress response and thiamine production.

Conclusions:

  • The study presents a new approach for high-throughput identification of genetic targets relevant to specific biosynthetic processes.
  • This method enhances the understanding of microbial metabolism and its impact on chemical production.
  • The findings are directly relevant to optimizing microbial cell factories.