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A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
Published on: October 4, 2019
Enhancing isoprenoid production through systematically assembling and modulating efflux pumps in Escherichia coli
Jian-Feng Wang1, Zhi-Qiang Xiong, Shi-Yuan Li
1Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, People's Republic of China.
Applied Microbiology and Biotechnology
|July 19, 2013
Summary
Engineering efflux pumps in Escherichia coli enhances the production of valuable compounds like amorphadiene and kaurene. Overexpressing specific pump components significantly boosted yields, demonstrating a promising strategy for microbial cell factories.
Area of Science:
- Synthetic Biology
- Metabolic Engineering
- Microbial Biotechnology
Background:
- Improving the yield of heterologous compounds in microbial cell factories is crucial for industrial applications.
- Efflux pumps facilitate the export of substances from microbial cells, enhancing product recovery and cellular tolerance.
- Understanding efflux pump mechanisms is key to optimizing compound production.
Purpose of the Study:
- To enhance the cellular exportation and production yield of hydrophobic isoprenoids (amorphadiene and kaurene) in Escherichia coli.
- To systematically engineer and modulate efflux pump systems (AcrAB-TolC, MdtEF-TolC, MexAB-OprM) for improved isoprenoid production.
Main Methods:
- Overexpression, assembly, and fine-tuning of native (E. coli) and heterologous (Pseudomonas aeruginosa) tripartite efflux pumps.
- Construction of various component combinations and modulation of key component copy numbers (AcrB, TolC).
- Quantification of specific yields for amorphadiene and kaurene production in engineered E. coli strains.
Main Results:
- Overexpression of AcrB and TolC components individually enhanced amorphadiene and kaurene yields by 31-37%.
- The heterologous MexB component significantly improved kaurene production by 70%.
- Specific combinations, such as TolC-TolC-AcrB and AcrA-TolC-AcrB, led to substantial yield increases of 118% and 104% respectively.
Conclusions:
- Systematic engineering and fine-tuning of efflux pumps, including component combinations and copy number modulation, is an effective strategy.
- Different efflux pump components exhibit varied effects on the production of specific isoprenoids.
- This approach offers a viable method for improving the production of diverse heterologous compounds in microbial hosts like E. coli.
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