title: Synthesis of methyl 5-O-trans-feruloyl-alpha-L-arabinofuranoside and its use as a substrate to assess feruloyl esterase activity.
abstract: A synthetic scheme was developed for the production of methyl 5-O-trans-feruloyl-alpha-L-arabinofuranoside (FA-Ara) in gram quantities. This molecule accurately models the chemical attachment of ferulic acid to polysaccharides found in cell walls of plants in the Gramineae family. It is therefore a realistic substrate that can be used to monitor feruloyl esterase activity. Ultraviolet spectral analysis indicated that FA-Ara has an absorption maximum distinct from the hydrolytic product, ferulic acid (FA), over a wide range of solution pH values. The log molar extinction coefficient ranges from 4.16 to 4.36 for FA-Ara and 4.16 to 4.33 for FA depending upon the pH of the buffered solution. Consequently a convenient spectrophotometric assay can be utilized to monitor esterase activity. Three different methods were developed for using this model substrate to assess esterase activity, including thin-layer chromatography, a spectrophotometric assay, and the use of high-performance liquid chromatography.
authors.affiliation: USDA-Agricultural Research Service, U.S. Dairy Forage Research Center, Madison, Wisconsin.
authors.forename: R D
authors.initials: RD
authors.lastname: Hatfield
authors.forename: R F
authors.initials: RF
authors.lastname: Helm
authors.forename: J
authors.initials: J
authors.lastname: Ralph
mesh_terms: D001089:Arabinose / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003373:Coumaric Acids / Q000138:chemical synthesis*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D013053:Spectrophotometry; D013379:Substrate Specificity
chemical_list: D003373:Coumaric Acids; C070193:methyl 5-O-feruloylarabinofuranoside; D001089:Arabinose; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase