title: Synthesis of 4-nitrophenyl caffeate and its use in assays of caffeoyl esterases.
abstract: We have prepared 4-nitrophenyl caffeate by a combination of standard procedures of organic synthesis and enzymatic deacetylation. Based on hydrolysis of 4-nitrophenyl caffeate, a convenient spectrophotometric assay was developed for specific monitoring of caffeoyl esterase. The method is fast and easy to perform, and it requires no expensive equipment. Its reliability was tested on eight enzyme preparations comprising various combinations of caffeoyl, feruloyl, and acetyl esterase as well as protease activities.
authors.affiliation: Institute of Chemistry, Center for Glycomics, Slovak Academy of Sciences, SK-845 38 Bratislava, Slovakia.
authors.forename: Andrej
authors.initials: A
authors.lastname: Chyba
authors.forename: Vladimír
authors.initials: V
authors.lastname: Mastihuba
authors.forename: Mária
authors.initials: M
authors.lastname: Mastihubová
mesh_terms: D055162:Biocatalysis; D002109:Caffeic Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D057075:Enzyme Assays; D009596:Nitrophenols / Q000737:chemistry*; D013053:Spectrophotometry; D013379:Substrate Specificity
chemical_list: D002109:Caffeic Acids; D009596:Nitrophenols; C024719:4-nitrophenyl; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase; C040048:caffeic acid
keywords: 4-Nitrophenyl caffeate; Caffeoyl esterase; HPLC assay; Spectrophotometric assay