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De novo production of resveratrol from glucose or ethanol by engineered Saccharomyces cerevisiae

M. Li ; K. R. Kildegaard ; Yun Chen (Institutionen för biologi och bioteknik, Systembiologi) ; A. Rodriguez ; I. Borodina ; Jens B. Nielsen (Institutionen för biologi och bioteknik, Systembiologi)
Metabolic Engineering (1096-7176). Vol. 32 (2015), p. 1-11.
[Artikel, refereegranskad vetenskaplig]

Resveratrol is a natural antioxidant compound, used as food supplement and cosmetic ingredient. Microbial production of resveratrol has until now been achieved by supplementation of expensive substrates, p-coumaric acid or aromatic amino acids. Here we engineered the yeast Saccharomyces cerevisiae to produce resveratrol directly from glucose or ethanol via tyrosine intermediate. First we introduced the biosynthetic pathway, consisting of tyrosine ammonia-lyase from Herpetosiphon aurantiacus, 4-coumaryl-CoA ligase from Arabidopsis thaliana and resveratrol synthase from Vitis vinifera, and obtained 2.73±0.05mgL-1 resveratrol from glucose. Then we over-expressed feedback-insensitive alleles of ARO4 encoding 3-deoxy-D-arabino-heptulosonate-7-phosphate and ARO7 encoding chorismate mutase, resulting in production of 4.85±0.31mgL-1 resveratrol from glucose as the sole carbon source. Next we improved the supply of the precursor malonyl-CoA by over-expressing a post-translational de-regulated version of the acetyl-CoA carboxylase encoding gene ACC1; this strategy further increased resveratrol production to 6.39±0.03mgL-1. Subsequently, we improved the strain by performing multiple-integration of pathway genes resulting in resveratrol production of 235.57±7.00mgL-1. Finally, fed-batch fermentation of the final strain with glucose or ethanol as carbon source resulted in a resveratrol titer of 415.65 and 531.41mgL-1, respectively.

Nyckelord: Metabolic engineering, Resveratrol, Saccharomyces cerevisiae, Tyrosine

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Denna post skapades 2015-10-08. Senast ändrad 2017-01-17.
CPL Pubid: 223810


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Institutionen för biologi och bioteknik, Systembiologi


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