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Oxidation-reduction or redox reactions involve the transfer of electrons from one molecule or atom to another. When an atom gains an electron, another atom must lose an electron, meaning oxidation and reduction must occur together. Since the redox occurs in pairs, the atom that gets oxidized is also called the reducing agent or reductant, and the atom that is reduced is also called the oxidizing agent or oxidant. A straightforward way to remember the definitions of oxidation and reduction is...
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Redox reactions are vital biochemical processes that underpin energy metabolism in cells. These reactions involve the transfer of electrons between molecules, occurring in tandem as oxidation and reduction. Oxidation refers to the loss of electrons, while reduction denotes their gain. This coupling ensures the seamless flow of electrons through metabolic pathways. For example, in bacterial metabolism, glucose undergoes oxidation to carbon dioxide, while oxygen is simultaneously reduced to...
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Light-Controlled Fermentations for Microbial Chemical and Protein Production
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Development of redox potential-driven fermentation process for recombinant protein expression.

Jingjie Guo1, Yixuan Wu1, Takuji Tanaka2

  • 1Department of Chemical and Biological Engineering, University of Saskatchewan, Saskatoon, Canada.

Biotechnology Letters
|October 16, 2020
PubMed
Summary
This summary is machine-generated.

This study developed a redox potential-driven fermentation process to optimize recombinant protein production in E. coli. The process accurately guides inducer addition and cell harvesting, improving prolinase activity.

Keywords:
Bioprocess developmentFermentationFermentation redox potentialRecombinant protein

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Area of Science:

  • Biotechnology
  • Microbial Fermentation
  • Protein Expression

Background:

  • Optimizing recombinant protein expression in microbial systems is crucial for industrial applications.
  • Traditional methods for guiding fermentation processes, such as optical density measurements, can be imprecise.
  • Redox potential offers a dynamic indicator of cellular metabolic state during fermentation.

Purpose of the Study:

  • To develop and evaluate a novel redox potential-driven fermentation strategy for guiding recombinant protein expression.
  • To establish clear indicators within the redox potential profile for critical process steps like induction and harvesting.
  • To compare the efficiency of the redox potential-guided process with traditional optical density-based methods.

Main Methods:

  • A redox potential-driven fermentation strategy was developed, maintaining dissolved oxygen at a specific level.
  • Recombinant E. coli strains expressing prolinase were cultivated using the developed process.
  • Oxidation-reduction potential (ORP) profiles were continuously monitored to identify key inflection points.
  • The timing of inducer addition and cell harvesting was guided by distinct ORP valleys.

Main Results:

  • Two distinct oxidation-reduction potential (ORP) valleys were identified during the fermentation of prolinase-expressing E. coli.
  • The first ORP valley accurately indicated the optimal time for adding the inducing agent.
  • The second ORP valley effectively guided the optimal timing for cell harvesting, leading to higher enzyme activity.
  • The developed process resulted in a prolinase activity of 0.172 μmol/mg/min, surpassing the 0.154 μmol/mg/min achieved with optical density-guided methods.

Conclusions:

  • The developed redox potential-driven fermentation process provides a more precise method for guiding recombinant protein expression.
  • The identified ORP valleys serve as reliable indicators for critical process control points.
  • The process demonstrates potential for further development into an automated fermentation system, enhancing efficiency and reproducibility.