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Improving formaldehyde consumption drives methanol assimilation in engineered E. coli
Benjamin M Woolston1, Jason R King1,2, Michael Reiter1
1Department of Chemical Engineering, Massachusetts Institute of Technology, 25 Ames Street, MIT 56-469C, Cambridge, MA, 02139, USA.
Nature Communications
|June 21, 2018
Summary
Methanol is a promising bio-based feedstock, but Escherichia coli struggles to metabolize it. This study identifies key bottlenecks in methanol assimilation and proposes solutions for improved bio-production.
Area of Science:
- Biotechnology
- Metabolic Engineering
- Synthetic Biology
Background:
- Methanol is a key feedstock for the bio-based economy due to economic and supply factors.
- Engineering Escherichia coli for efficient methanol metabolism presents significant challenges and bottlenecks.
Purpose of the Study:
- To systematically analyze and identify bottlenecks in methanol assimilation pathways in Escherichia coli.
- To develop strategies to overcome these limitations and enhance methanol utilization.
Main Methods:
- Systematic analysis of potential pathway bottlenecks in methanol assimilation.
- Activation of the sedoheptulose bisphosphatase variant of the ribulose monophosphate pathway.
- Utilizing kinetic isotope effects with deuterated methanol to probe pathway flux limitations.
Main Results:
- Ribulose 5-phosphate regeneration is insufficient for methanol assimilation in E. coli.
- Activating a specific pathway variant successfully addressed regeneration limitations.
- Methanol dehydrogenase was identified as the rate-limiting enzyme for overall pathway flux.
- NADH was found to be a potent kinetic inhibitor of methanol dehydrogenase.
Conclusions:
- Overcoming identified bottlenecks is crucial for efficient methanol utilization in engineered E. coli.
- Strategies include pathway optimization and addressing enzyme inhibition.
- Chemical biology approaches are valuable for advancing metabolic engineering efforts.
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