|Cote, Gregory - Greg|
Submitted to: Analytical Biochemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/12/2002
Publication Date: 10/1/2002
Citation: MASTIHUBA, V., KREMNICKY, L., MASTIHUBOVA, M., WILLETT, J.L., COTE, G.L. A SPECTROPHOTOMETRIC ASSAY FOR FERULOYL ESTERASES. ANALYTICAL BIOCHEMISTRY. 2002. V. 309. P. 96-101. Interpretive Summary: The enzyme FAE helps break down certain types of chemical bonds in plant cell walls. This process is important in digestion of forage plants by ruminants such as sheep and cattle. The enzyme is also of interest in the modification of plant matter for extraction of natural products. This paper describes a new method of analyzing the activity of FAE enzymes. A key step is the use of a new compound recently synthesized by our group which involves use of a biocatalyst, rather than harsh chemical methods. This research provides a new tool for scientists styding this important enzyme, enabling more productive research in this area.
Technical Abstract: 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.