Automated determination of microbial peroxidase activity in fermentation samples using hydrogen peroxide as the substrate and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) as the electron donor in a flow injection system

Abstract

An automated flow injection method has been developed for the determination of microbial peroxidase activity. The substrate used was hydrogen peroxide and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate)(ABTS) was used as the electron donor. In the presence of hydrogen peroxide, peroxidase catalyses the dehydrogenation of ABTS, resulting in the formation of a resonance-stabilized radical cation of ABTS. The green–blue colour formed, recorded at 418 nm, is taken as a measure of the peroxidase activity. The general technical conditions and the general enzymic kinetics have been optimized. Conditions for activation and stabilization of the enzyme were found, e.g., ammonium sulfate acts as a peroxidase activator. The resulting method has a good precision, sensitivity and speed.