Using unnatural protein fusions to engineer resveratrol biosynthesis in yeast and Mammalian cells

J Am Chem Soc. 2006 Oct 11;128(40):13030-1. doi: 10.1021/ja0622094.

Abstract

Resveratrol is a naturally occurring defense compound produced by a limited number of plants in response to stresses. Besides cardiovascular benefits, this health-promoting compound has been reported to extend life spans in yeasts, flies, worms, and fish. To biosynthesize resveratrol de novo, tyrosine ammonia lyase (TAL), 4-coumarate CoA-ligase (4CL), and stilbene synthase (STS) were isolated from Rhodobacter sphaeroides, Arabidopsis thaliana, and Vitis vinifera, respectively. Yeast cells expressing 4CL and STS produce resveratrol when fed with 4-coumaric acid, the substrate of 4CL. When a translational fusion protein joining 4CL and STS was used, yeast cells produced 15-fold more resveratrol than the cotransformed cells, suggesting that physical localization of 4CL and STS facilitate resveratrol production. When the resveratrol pathway was introduced into human HEK293 cells, de novo biosynthesis was detected, leading to intracellular accumulation of resveratrol. We successfully engineered an entire plant natural product pathway into a mammalian host.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acyltransferases / genetics*
  • Acyltransferases / metabolism
  • Ammonia-Lyases / genetics*
  • Ammonia-Lyases / metabolism
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism
  • Cell Line
  • Cloning, Molecular
  • Coenzyme A Ligases / genetics*
  • Coenzyme A Ligases / metabolism
  • Humans
  • Kidney / cytology
  • Protein Engineering / methods
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Resveratrol
  • Rhodobacter sphaeroides / enzymology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Stilbenes / metabolism*

Substances

  • Arabidopsis Proteins
  • Recombinant Fusion Proteins
  • Stilbenes
  • Acyltransferases
  • stilbene synthase
  • Ammonia-Lyases
  • L-tyrosine ammonia-lyase
  • 4CL1 protein, Arabidopsis
  • Coenzyme A Ligases
  • Resveratrol