Near UV difference spectroscopy application for ferulic acid esterases substrate specificity determination

Publication date: Available online 6 February 2017 Source:Journal of Molecular Catalysis B: Enzymatic Author(s): Gleb S. Dotsenko, Anna S. Dotsenko, Arkady P. Sinitsyn Ferulic acid esterases (FAEs) are promising enzymes for potential application in agricultural, food, paper, and pharmaceutical industries. Discovery and characterization of novel FAEs is still up to date. Depending on their substrate specificity towards four methyl hydroxycinnamates (methyl esters of ferulic, p-coumaric, caffeic, and sinapic acids), FAEs are classified into four subgroups, named A, B, C, and D types. In this work we investigate practical applicability of near UV difference spectroscopy for FAE’s substrate specificity determination. Difference spectroscopy is demonstrated to be a convenient and easily performed approach, enabling either single point or continuous determination of four methyl hydroxycinnamates hydrolysis rate at a concentration range 0.02-1.0mM. Difference maxima wavelengths and corresponding extinction coefficients are determined for free and methyl esterified ferulic, p-coumaric, caffeic, and sinapic acids. Graphical abstract

Source: Journal of Molecular Catalysis B: Enzymatic - February 6, 2017 Category: Biochemistry Source Type: research