title: Using unnatural protein fusions to engineer resveratrol biosynthesis in yeast and Mammalian cells.
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.
authors.affiliation: Donald Danforth Plant Science Center, St. Louis, Missouri 63132, USA.
authors.forename: Yansheng
authors.initials: Y
authors.lastname: Zhang
authors.forename: Song-Zhe
authors.initials: SZ
authors.lastname: Li
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.forename: Xiangqing
authors.initials: X
authors.lastname: Pan
authors.forename: Rebecca E
authors.initials: RE
authors.lastname: Cahoon
authors.forename: Jan G
authors.initials: JG
authors.lastname: Jaworski
authors.forename: Xuemin
authors.initials: X
authors.lastname: Wang
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Jez
authors.forename: Feng
authors.initials: F
authors.lastname: Chen
authors.forename: Oliver
authors.initials: O
authors.lastname: Yu
mesh_terms: D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism; D000642:Ammonia-Lyases / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D003066:Coenzyme A Ligases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D000077185:Resveratrol; D012242:Rhodobacter sphaeroides / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D013267:Stilbenes / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D011993:Recombinant Fusion Proteins; D013267:Stilbenes; D000217:Acyltransferases; C022674:stilbene synthase; D000642:Ammonia-Lyases; C023208:L-tyrosine ammonia-lyase; C511052:4CL1 protein, Arabidopsis; D003066:Coenzyme A Ligases; D000077185:Resveratrol