Metabolically engineered cells for the production of resveratrol or an oligomeric or glycosidically-bound derivative thereof

Patent metadata

Field	Value
Publication number	US-9404129-B2
Application number	US-201414514718-A
Country	US
Kind code	B2
Filing date	Oct 15, 2014
Priority date	Feb 22, 2005
Publication date	Aug 2, 2016
Grant date	Aug 2, 2016

Abstract

A recombinant micro-organism producing resveratrol by a pathway in which phenylalanine ammonia lyase (PAL) produces trans-cinnamic acid from phenylalanine, cinnamate 4-hydroxylase (C4H) produces 4-coumaric acid from said trans-cinnamic acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA, or in which L-phenylalanine- or tyrosine-ammonia lyase (PAL/TAL) produces 4-coumaric acid, 4-coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4-coumaroyl CoA. The micro-organism may be a yeast, fungus or bacterium including Saccharomyces cerevisiae, E. coli, Lactococcus lactis, Aspergillus niger , or Aspergillus oryzae.

First claim

The invention claimed is:

1. A method for producing resveratrol or an oligomeric or glycosidically-bound derivative thereof comprising:
a) cultivating a recombinant micro-organism comprising an engineered operative metabolic pathway producing resveratrol or an oligomeric or glycosidically-bound derivative thereof in a culture media comprising a carbon substrate from which the micro-organism can produce resveratrol, wherein the culture media does not require an external source of coumaric acid, and wherein the operative metabolic pathway produces:
i) 4-coumaric acid from L-phenylalanine catalysed by a phenylalanine ammonia lyase (PAL) and a cinnamate 4-hydroxylase (C4H) expressed in the micro-organism or from tyrosine catalysed by a phenylalanine ammonia lyase (PAL) or a tyrosine ammonia lyase (TAL) expressed in said micro-organism; and
ii) 4-coumaroyl-CoA from 4-coumaric acid catalysed by a 4-coumarate-CoA ligase (4CL) expressed in said micro-organism; and
iii) resveratrol is produced from the 4-coumaroyl-CoA by a resveratrol synthase expressed in the micro-organism; and

b) recovering the resveratrol or the oligomeric or glycosidically-bound derivative thereof from the culture media.

2. The method of claim 1 , wherein the micro-organism is fungi.

3. The method of claim 2 , wherein the fungus is yeast.

4. The method of claim 3 , wherein the yeast is from the genus Saccharomyces.

5. The method of claim 1 , wherein the carbon substrate is a fermentable carbon substrate.

6. The method of claim 5 , wherein the fermentable carbon substrate is monosaccharides, oligosaccharides or polysaccharides.

7. The method of claim 5 , wherein the fermentable carbon substrate is glucose, fructose, galactose, xylose, arabinose, mannose, sucrose, lactose, erythrose, threose or ribose.

8. The method of claim 1 , wherein the carbon substrate is a non-fermentable carbon substrate.

9. The method of claim 8 , wherein the non-fermentable carbon substrate is ethanol, acetate, glycerol and lactate.

10. The method of claim 1 , wherein the resveratrol or the oligomeric or glycosidically-bound derivative thereof recovered from the culture media comprises a nutraceutical in a dairy product or a beverage.

11. The method of claim 1 , wherein at least 0.44-0.53 ug of resveratrol per gram dry weight of the recombinant microorganism is produced.

Assignees

• Evolva Sa

Inventors

• Katz Michael
• Smits Hans Peter
• Forster Jochen
• Nielsen Jens Bredel

Classifications

• C12P7/42

  Hydroxy-carboxylic acids · CPC title

• C12N9/0042

  NADPH-cytochrome P450 reductase (1.6.2.4) · CPC title

• C12N9/1037

  Naringenin-chalcone synthase (2.3.1.74), i.e. chalcone synthase · CPC title

• C12P7/22 Primary

  aromatic · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title