title: Metabolic engineering for resveratrol derivative biosynthesis in Escherichia coli.
abstract: We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.
authors.forename: Yu Jeong
authors.initials: YJ
authors.lastname: Jeong
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.
authors.forename: Su Gyeong
authors.initials: SG
authors.lastname: Woo
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.|Department of Bioscience and Biotechnology, Chungnam National University, Daejeon 305-764, Korea.
authors.forename: Chul Han
authors.initials: CH
authors.lastname: An
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.
authors.forename: Hyung Jae
authors.initials: HJ
authors.lastname: Jeong
authors.affiliation: Chemical Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Ochang 363-883, Korea.
authors.forename: Young-Soo
authors.initials: YS
authors.lastname: Hong
authors.affiliation: Department of Food Science and Technology and Functional Food Research Center, Chonnam National University, Gwangju 500-757, Korea.
authors.forename: Young-Min
authors.initials: YM
authors.lastname: Kim
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.
authors.forename: Young Bae
authors.initials: YB
authors.lastname: Ryu
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.
authors.forename: Mun-Chual
authors.initials: MC
authors.lastname: Rho
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.
authors.forename: Woo Song
authors.initials: WS
authors.lastname: Lee
authors.affiliation: Eco-friendly Bio-Material Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 580-185, Korea.
authors.forename: Cha Young
authors.initials: CY
authors.lastname: Kim
mesh_terms: D000217:Acyltransferases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D000077185:Resveratrol; D013267:Stilbenes / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D013267:Stilbenes; C522997:pinostilbene; D008780:Methyltransferases; D000217:Acyltransferases; C022674:stilbene synthase; D000077185:Resveratrol
keywords: metabolic engineering; pinostilbene; resveratrol; resveratrol O-methyltransferase; stilbene synthase