Continuous kinetic assay of arylsulfatases with new chromogenic and fluorogenic substrates

Arylsulfatases are determined, evenin weakly acidic solution, by a direct and continuous kinetic method using new coumarin-derived sulfates as substrates. After enzymatic hydrolysis, the substrate dissociates to form intensely colored and strongly fluorescent phenolates, with absorption maxima ranging from 383 to 497 nm, and fluorescence emission maxima between 470 and 577 nm. Rates of enzymatic hydrolysis, optimum pH values and detection limits of arylsulfatase from Aerobacter aerogenes and Patella vulgata are determined.