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Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade
Published on: August 14, 2019
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Enhancing Flavan-3-ol Biosynthesis in Saccharomyces cerevisiae
Chengcheng Sun1,2,3,4, Guangjian Li1,2,4, Hongbiao Li1,2,3,4
1National Engineering Laboratory for Cereal Fermentation Technology (NELCF), Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
Journal of Agricultural and Food Chemistry
|October 25, 2021
Summary
Researchers engineered yeast to produce flavan-3-ols, afzelechin (AFZ) and catechin (CAT), from glucose. Optimized metabolic pathways and enzyme combinations achieved high yields of these beneficial compounds.
Area of Science:
- Metabolic Engineering
- Synthetic Biology
- Biotechnology
Background:
- Flavan-3-ols are beneficial flavonoids with potential health applications.
- Current methods for flavan-3-ol production are limited.
- Engineering microorganisms offers a sustainable production route.
Purpose of the Study:
- To engineer Saccharomyces cerevisiae for de novo biosynthesis of afzelechin (AFZ) and catechin (CAT).
- To optimize metabolic pathways and enzyme activities for enhanced flavan-3-ol production.
Main Methods:
- Genetic engineering of Saccharomyces cerevisiae strain E32.
- Introduction of key enzymes: flavonoid 3-hydroxylase, flavonoid 3'-hydroxylase, dihydroflavonol 4-reductase (DFR), and leucoanthocyanidin reductase (LAR).
- Optimization of enzyme combinations, linkers, promoters, and NADPH supply.
Main Results:
- Achieved de novo production of AFZ and CAT in engineered yeast.
- Identified optimal enzyme combinations (FaDFR and VvLAR) and linkers ((GGGGS)2 for AFZ, (EAAAK)2 for CAT).
- Reached production levels of 500.5 mg/L AFZ and 321.3 mg/L CAT in a 5 L bioreactor after 90 h fermentation.
Conclusions:
- Demonstrated successful engineering of yeast for high-yield flavan-3-ol production.
- Developed strategies applicable for efficient microbial production of various flavan-3-ols.
- This approach provides a scalable and sustainable method for producing valuable flavan-3-ols.
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