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Light-Controlled Fermentations for Microbial Chemical and Protein Production

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Published on: March 22, 2022

Characterizing and utilizing oxygen-dependent promoters for efficient dynamic metabolic engineering.

Julian Wichmann¹, Gerrich Behrendt¹, Simon Boecker¹

¹Analysis and Redesign of Biological Networks, Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106, Magdeburg, Germany.

Metabolic Engineering

|April 13, 2023

Summary

Researchers characterized 15 oxygen-depleted promoters in Escherichia coli for dynamic metabolic engineering. Six promoters were identified as ideal for enhancing microbial productivity through controlled gene expression, significantly boosting glucose uptake rates.

Keywords:

Bioprocess design Dynamic metabolic engineering Enforced ATP wasting Escherichia coli Lactate Oxygen-responsive promoters Two-stage processes

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Area of Science:

Synthetic Biology
Metabolic Engineering
Microbial Fermentation

Background:

Promoters regulate gene expression in response to cellular signals, crucial for dynamic metabolic engineering in fermentation.
Dissolved oxygen is a key signal, as production often occurs under anaerobic conditions.
A comprehensive comparison of oxygen-dependent promoters in Escherichia coli is lacking.

Purpose of the Study:

To systematically test and characterize 15 promoter candidates induced by oxygen depletion in Escherichia coli.
To identify suitable promoters for dynamic metabolic engineering applications.
To demonstrate the utility of selected promoters in optimizing microbial productivity.

Main Methods:

Developed a microtiter plate-level screening assay using an oxygen-independent fluorescent protein.
Employed flow cytometry for verification of promoter activity.
Assessed promoter performance in dynamic induction of enforced ATP wasting and optimization of lactate production.

Main Results:

Identified six promoters (nar-strong, nar-medium, nar-weak, nirB-m, yfiD-m, fnrF8) suitable for dynamic metabolic engineering.
Demonstrated that selected promoters exhibit tight regulation under aerobic conditions and high expression under anaerobiosis.
Achieved unprecedented specific glucose uptake rates by dynamically enforcing ATP wasting using selected promoters.

Conclusions:

The characterized promoters provide valuable tools for implementing oxygen-inducible dynamic metabolic engineering strategies.
Dynamic induction of ATP wasting using selected promoters significantly enhances microbial productivity.
Optimized a two-stage lactate production process, showcasing the practical application of these promoters.