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Optogenetic Amplification Circuits for Light-Induced Metabolic Control.

Evan M Zhao1, Makoto A Lalwani1, Jhong-Min Chen1

  • 1Department of Chemical and Biological Engineering, Princeton University, Princeton, New Jersey 08544, United States.

ACS Synthetic Biology
|April 9, 2021
PubMed
Summary

New optogenetic circuits (OptoAMP) amplify light responses in microbial fermentations, overcoming light penetration issues. This enables dynamic metabolic control for improved chemical production, even in large bioreactors.

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Area of Science:

  • Synthetic Biology
  • Metabolic Engineering
  • Biotechnology

Background:

  • Dynamic control of microbial metabolism enhances chemical production during fermentation.
  • Optogenetics offers tunable, reversible control using light, but faces challenges with light penetration in dense cell cultures and large bioreactors.

Purpose of the Study:

  • To develop novel optogenetic circuits (OptoAMP) that amplify transcriptional responses to light.
  • To address limitations of light penetration in high-cell-density fermentations.

Main Methods:

  • Designed and characterized OptoAMP circuits, comparing them to a basal circuit (OptoEXP).
  • Evaluated circuit performance in bioreactors up to 5 L, assessing induction levels, light sensitivity, and homogeneity.
  • Applied OptoAMP circuits to control engineered metabolic pathways in three-phase fermentations for chemical production.

Main Results:

  • OptoAMP circuits provided up to a 23-fold amplification of transcriptional response compared to OptoEXP.
  • Achieved up to 41-fold induction between dark and light conditions with efficient activation at low light duty cycles (∼1%).
  • Demonstrated strong, homogeneous light induction in large bioreactors (≥5 L) even at high cell densities ( > 40 OD600).

Conclusions:

  • OptoAMP circuits significantly enhance the applicability of optogenetics in metabolic engineering.
  • These circuits enable precise, dynamic control of microbial metabolism for improved production of valuable chemicals like lactic acid, isobutanol, and naringenin.