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Quantitative metabolomics for dynamic metabolic engineering using stable isotope labeled internal standards mixture
Yuki Soma1, Masatomo Takahashi1, Yuri Fujiwara1
1Division of Metabolomics, Research Center for Transomics Medicine, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Journal of Bioscience and Bioengineering
|October 8, 2021
Summary
Dynamic metabolic engineering optimizes microbial fermentation for sustainable chemical production. Quantitative metabolome analysis identified bottlenecks in engineered E. coli for improved gamma-aminobutyrate (GABA) fermentation.
Area of Science:
- Synthetic Biology
- Metabolic Engineering
- Biotechnology
Background:
- Microbial production of chemicals and fuels from renewables offers an alternative to fossil fuels.
- Metabolic engineering enables pathway optimization in microorganisms.
- Dynamic metabolic engineering, using synthetic biology, is an advanced strategy for optimizing fermentation.
Purpose of the Study:
- To investigate the impact of dynamic metabolic flux regulation on engineered Escherichia coli for gamma-aminobutyrate (GABA) production.
- To identify bottlenecks in GABA fermentation using quantitative metabolome analysis.
Main Methods:
- Quantitative metabolome analysis using mass spectrometry.
- Stable isotope dilution method (SIDM) with a stable isotope-labeled internal standard mixture (SILIS).
- Engineering of Escherichia coli for dynamic metabolic flux regulation.
Main Results:
- Identified multiple candidate bottlenecks limiting GABA production.
- Demonstrated the utility of dynamic metabolic engineering for optimizing fermentation processes.
- Quantitative metabolome analysis revealed specific metabolic reactions requiring further engineering.
Conclusions:
- Dynamic metabolic engineering strategies require iterative evaluation and redesign for practical application.
- Further engineering of specific metabolic reactions is necessary for enhanced direct GABA fermentation.
- Quantitative metabolomics is crucial for understanding and improving engineered microbial systems.
