title: Genetic engineering approach for the production of rhamnosyl and allosyl flavonoids from Escherichia coli.
abstract: The main functions of glycosylation are stabilization, detoxification and solubilization of substrates and products. To produce glycosylated products, Escherichia coli was engineered by overexpression of TDP-L-rhamnose and TDP-6-deoxy-D-allose biosynthetic gene clusters, and flavonoids were glycosylated by the overexpression of the glycosyltransferase gene from Arabidopsis thaliana. For the glycosylation, these flavonoids (quercetin and kaempferol) were exogenously fed to the host in a biotransformation system. The products were isolated, analyzed and confirmed by HPLC, LC/MS, and ESI-MS/MS analyses. Several conditions (arabinose, IPTG concentration, OD(600), substrate concentration, incubation time) were optimized to increase the production level. We successfully isolated approximately 24 mg/L 3-O-rhamnosyl quercetin and 12.9 mg/L 3-O-rhamnosyl kaempferol upon feeding of 0.2 mM of the respective flavonoids and were also able to isolate 3-O-allosyl quercetin. Thus, this study reveals a method that might be useful for the biosynthesis of rhamnosyl and allosyl flavonoids and for the glycosylation of related compounds.
authors.affiliation: Institute of Biomolecule Reconstruction, Department of Pharmaceutical Engineering, Sun Moon University, Tangjeong-myeon, Asansi, Chungnam, Korea.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Simkhada
authors.forename: Hei Chan
authors.initials: HC
authors.lastname: Lee
authors.forename: Jae Kyung
authors.initials: JK
authors.lastname: Sohng
mesh_terms: D004926:Escherichia coli / Q000502:physiology*; D005419:Flavonoids / Q000378:metabolism*; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D009702:Nucleoside Diphosphate Sugars / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D012210:Rhamnose / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D012431:Rutin / Q000378:metabolism*; D013942:Thymine Nucleotides / Q000235:genetics / Q000378:metabolism*
chemical_list: D005419:Flavonoids; D017127:Glycation End Products, Advanced; D009702:Nucleoside Diphosphate Sugars; D013942:Thymine Nucleotides; C119260:dTDP-4-ketorhamnose; C518397:deoxythymidine diphosphate-6-deoxyallose; D012431:Rutin; D012210:Rhamnose