title: Statistically designed enzymatic hydrolysis for optimized production of icariside II as a novel melanogenesis inhibitor.
abstract: Three kinds of prenylated flavonols, icariside I, icariside II, and icaritin, were isolated from an icariin hydrolysate and their effects on melanogenesis evaluated based on mushroom tyrosinase inhibition and quantifying the melanin contents in melanocytes. Although none of the compounds had an effect on tyrosinase activity, icariside II and icaritin both effectively inhibited the melanin contents with an IC50 of 10.53 and 11.13 MM, respectively. Whereas icariside II was obtained from a reaction with beta-glucosidase and cellulase, the icariin was not completely converted into icariside II. Thus, for the high-purity production of icariside II, the reaction was optimized using the response surface methodology, where an enzyme concentration of 5.0 mg/ml, pH 7, 37.5 degrees C;, and 8 h reaction time were selected as the central conditions for the central composite design (CCD) for the enzymatic hydrolysis of icariin into icariside II using cellulase. Empirical models were developed to describe the relationships between the operating factors and the response (icariside II yield). A statistical analysis indicated that all four factors had a significant effect (p<0.01) on the icariside II production. The coefficient of determination (R2) was good for the model (0.9853), and the optimum production conditions for icariside II was an enzyme concentration of 7.5 mg/ml, pH 5, 50 degrees C, and 12 h reaction time. A good agreement between the predicted and experimental data under the designed optimal conditions confirmed the usefulness of the model. A laboratory pilot scale was also successful.
authors.affiliation: Skin Research Institute, Amorepacific R&D Center, Yongin 449-729, Korea. superbody@amorepacific.com
authors.forename: Jun-Seong
authors.initials: JS
authors.lastname: Park
authors.forename: Hye Yoon
authors.initials: HY
authors.lastname: Park
authors.forename: Ho-Sik
authors.initials: HS
authors.lastname: Rho
authors.forename: Soomi
authors.initials: S
authors.lastname: Ahn
authors.forename: Duck Hee
authors.initials: DH
authors.lastname: Kim
authors.forename: Ih Seop
authors.initials: IS
authors.lastname: Chang
mesh_terms: D000818:Animals; D001709:Biotechnology; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D034181:Epimedium / Q000737:chemistry*; D047309:Flavones / Q000378:metabolism / Q000494:pharmacology; D005419:Flavonoids / Q000378:metabolism* / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D013058:Mass Spectrometry; D008543:Melanins / Q000037:antagonists & inhibitors*; D008544:Melanocytes; D051379:Mice; D008810:Mice, Inbred C57BL; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D014468:Umbelliferones / Q000378:metabolism / Q000494:pharmacology
chemical_list: D047309:Flavones; D005419:Flavonoids; D008543:Melanins; D014468:Umbelliferones; C526898:icariside I; C060988:baohuoside I; D014442:Monophenol Monooxygenase; C499403:icaritin; C056599:icariin