title: Preparation and properties of rutin-hydrolyzing enzyme from tartary buckwheat seeds.
abstract: A rutin hydrolyzing enzyme (RHE) was isolated from Fagopyrum tataricum Moench seeds by using ammonium sulphate fractionation, anion exchange and size exclusion chromatography. The purified RHE has an apparent molecular weight of about 70kDa determined by SDS-PAGE, with an isoelectric point (pI) (determined by isoelectric focusing) of 6.7. RHE has a specific catalytic activity toward rutin when incubated together with rutin at 37°C for 30min in the presence of 20% ethanol, and its Km value for rutin is 1.04×10(-3)M. The RHE catalytic product analyzed by HPLC displayed high similarity with quercetin and this is confirmed by (1)H NMR spectroscopy and LC-ESI-MS/MS, suggesting that the RHE hydrolysis product is quercetin. These results suggest that the RHE from tartary buckwheat seeds is a specific rutin-hydrolyzing enzyme, providing a new enzymatic preparation method for quercetin.
authors.affiliation: Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, PR China.
authors.forename: Xiao-Dong
authors.initials: XD
authors.lastname: Cui
authors.affiliation: Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, PR China. Electronic address: zhwang@sxu.edu.cn.
authors.forename: Zhuan-Hua
authors.initials: ZH
authors.lastname: Wang
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D019612:Fagopyrum / Q000737:chemistry*; D006868:Hydrolysis; D011794:Quercetin / Q000032:analysis / Q000737:chemistry*; D012431:Rutin / Q000032:analysis / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry*
chemical_list: D012431:Rutin; D011794:Quercetin
keywords: Fagopyrum tataricum Moench; Quercetin; Rutin; Rutin-hydrolyzing enzyme