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A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
Published on: October 4, 2019
11.9K
Combining genotype improvement and statistical media optimization for isoprenoid production in E. coli
Congqiang Zhang1, Xixian Chen, Ruiyang Zou
1Chemical and Pharmaceutical Engineering, Singapore-MIT Alliance, Singapore, Singapore.
Plos One
|October 15, 2013
Summary
Metabolic engineering of E. coli by deleting PTS genes enhanced phosphoenolpyruvate availability, significantly boosting isoprenoid production. This strategy achieved a seven-fold increase in lycopene yield, demonstrating a new method for sustainable natural product biosynthesis.
Area of Science:
- Metabolic Engineering
- Synthetic Biology
- Biotechnology
Background:
- Isoprenoids are high-value compounds with increasing demand.
- Metabolic engineering in microbes offers a sustainable production method.
- Optimizing metabolic precursors for the deoxyxylulose phosphate (DXP) pathway is crucial for high isoprenoid yields.
Purpose of the Study:
- To enhance isoprenoid production in E. coli by optimizing precursor availability.
- To investigate the effect of carbohydrate phosphotransferase system (PTS) deletion on DXP pathway precursors.
- To maximize lycopene production through medium and culture condition optimization.
Main Methods:
- Deletion of genes encoding the carbohydrate phosphotransferase system (PTS) in E. coli.
- Optimization of growth medium and culture conditions for microbial fermentation.
- Quantification of lycopene and amorpha-1,4-diene yields.
Main Results:
- Deletion of PTS genes enriched phosphoenolpyruvate, a key DXP pathway precursor.
- Optimized conditions led to a seven-fold higher lycopene yield in PTS mutants compared to wild type.
- A record yield of 20,000 µg/g dry cell weight for lycopene was achieved.
- The strategy successfully enhanced production of another isoprenoid, amorpha-1,4-diene.
Conclusions:
- Deleting PTS genes is an effective strategy to increase isoprenoid precursor availability.
- Optimized fermentation conditions further enhance microbial production of high-value isoprenoids.
- The developed metabolic engineering approaches are broadly applicable for optimizing the production of diverse isoprenoids.
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