Engineering yeast for high-level production of stilbenoid antioxidants.
Stilbenoids, including resveratrol and its methylated derivatives, are natural potent antioxidants, produced by some plants in trace amounts as defense compounds. Extraction of stilbenoids from natural sources is costly due to their low abundance and often limited availability of the plant. Here we engineered the yeast Saccharomyces cerevisiae for production of stilbenoids on a simple mineral medium typically used for industrial production. We applied a pull-push-block strain engineering strategy that included overexpression of the resveratrol biosynthesis pathway, optimization of the electron transfer to the cytochrome P450 monooxygenase, increase of the precursors supply, and decrease of the pathway intermediates degradation. Fed-batch fermentation of the final strain resulted in a final titer of 800 mg l-1resveratrol, which is by far the highest titer reported to date for production of resveratrol from glucose. We further integrated heterologous methyltransferases into the resveratrol platform strain and hereby demonstrated for the first time de novo biosynthesis of pinostilbene and pterostilbene, which have better stability and uptake in the human body, from glucose.
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2970 Hørsholm, Denmark.
Mingji
M
Li
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2970 Hørsholm, Denmark.
Konstantin
K
Schneider
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2970 Hørsholm, Denmark.
Mette
M
Kristensen
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2970 Hørsholm, Denmark.
Irina
I
Borodina
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2970 Hørsholm, Denmark.|Department of Biology and Biological Engineering, Chalmers University of Technology, SE-41296 Gothenburg, Sweden.|The Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE-41296 Gothenburg, Sweden.
Jens
J
Nielsen
D000975:Antioxidants / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics; D019149:Bioreactors; D053898:Biosynthetic Pathways; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics; D005285:Fermentation; D008316:Malonyl Coenzyme A / Q000378:metabolism; D060847:Metabolic Engineering; D010649:Phenylalanine / Q000378:metabolism; D011401:Promoter Regions, Genetic; D000077185:Resveratrol; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013267:Stilbenes / Q000378:metabolism*
D000975:Antioxidants; D029681:Arabidopsis Proteins; D013267:Stilbenes; C522997:pinostilbene; C107773:pterostilbene; D010649:Phenylalanine; D008316:Malonyl Coenzyme A; D003577:Cytochrome P-450 Enzyme System; D000077185:Resveratrol