A spectrophotometric assay for feruloyl esterases

• Published in: Analytical biochemistry Vol. 309; no. 1; pp. 96 - 101
• Main Authors: Mastihuba, Vladimı́r, Kremnický, Lubomı́r, Mastihubová, Mária, Willett, J.L, Côté, Gregory L
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.2002
• Subjects: 4-Nitrophenol; 4-Nitrophenyl ferulate; Assay method; carboxylic ester hydrolases; Carboxylic Ester Hydrolases - chemistry; Carboxylic Ester Hydrolases - metabolism; Chromatography, High Pressure Liquid - methods; Chromatography, Thin Layer; Endopeptidases - chemistry; Endopeptidases - metabolism; enzyme activity; enzyme substrates; Esterases - chemistry; Esterases - classification; Esterases - metabolism; ferulic acid; Feruloyl esterase; Hydrolysis; Kinetics; Lipase - chemistry; Lipase - metabolism; nitrophenols; Nitrophenols - analysis; Nitrophenols - metabolism; Octoxynol - chemistry; quantitative analysis; spectrophotometers; Spectrophotometry; Spectrophotometry, Ultraviolet - methods; Substrate Specificity; 4-Nitrophenyl ferulate; 4-Nitrophenol; Feruloyl esterase; Spectrophotometry; Assay method
• ISSN: 0003-2697; 1096-0309; 1096-0309
• DOI: 10.1016/S0003-2697(02)00241-5

We have developed a spectrophotometric assay for the quantitative determination of feruloyl esterase activity based on release of 4-nitrophenol from a novel substrate, 4-nitrophenyl ferulate in an emulsion of Triton X-100 in aqueous buffer solution. The release of 4-nitrophenol was linear with reaction time at an early stage of the reaction with various esterase preparations. The method proposed here is accurate, rapid, and easy to perform.