Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 69, Issue 43, Pages 12763-12772Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.1c04489
Keywords
catechin; afzelechin; flavan-3-ols; linker; metabolic engineering; Saccharomyces cerevisiae
Funding
- Foundation for Innovative Research Groups of the National Natural Science Foundation of China [32021005]
- National Science Fund for Excellent Young Scholars [21822806]
- National Natural Science Foundation of China [21908078]
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This study successfully achieved de novo production of two basic flavan-3-ols by engineering Saccharomyces cerevisiae, demonstrating the potential of engineering strategies in increasing yield.
Flavan-3-ols are a group of flavonoids that exert beneficial effects. This study aimed to enhance key metabolic processes related to flavan-3-ols biosynthesis. The engineered Saccharomyces cerevisiae strain E32 that produces naringenin from glucose was further engineered for de novo production of two basic flavan-3-ols, afzelechin (AFZ) and catechin (CAT). Through introduction of flavonoid 3-hydroxylase, flavonoid 3'-hydroxylase, dihydroflavonol 4-reductase (DFR), and leucoanthocyanidin reductase (LAR), de novo production of AFZ and CAT can be achieved. The combination of FaDFR from Fragaria x ananassa and VvLAR from Vitis vinifera was optimal. (GGGGS)(2) and (EAAAK)(2) linkers between DFR and LAR proved optimal for the production of AFZ and CAT, respectively. Optimization of promoters and the enhanced supply of NADPH further increased the production. By combining the best engineering strategies, the optimum strains produced 500.5 mg/L AFZ and 321.3 mg/L CAT, respectively, after fermentation for 90 h in a 5 L bioreactor. The strategies presented could be applied for a more efficient production of flavan-3-ols by various microorganisms.
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