Optimization of Peroxidase-Catalyzed Oxidation of Luminol in Reversed Micelles of Surfactants in Octane

Abstract

Luminol oxidation in the Aerosol OT (AOT) reversed micelles in octane catalyzed by horseradish peroxidase (HRP), or its conjugate with Cortisol (HRP-COR), was optimized. The chemiluminescence intensity during luminol oxidation was strongly dependent on the method of preparation of the reaction mixture and the addition of Triton X-45, cyclohexanol and the chemiluminescence “enhancer”, p-iodophenol, into the micellar system. Five procedures for the preparation of the reaction mixture were compared. The maximum chemiluminescence was observed in the micellar system containing all the reaction components excluding a biocatalyst, addition of which into the system started the reaction. Triton X-45, cyclohexanol or p-iodophenol added to the micellar system enhanced significantly the chemiluminescence intensity. The “enhancing” action of p-iodophenol in AOT reversed micelles was 10-fold less than in an aqueous medium.

Key Words:

• Peroxidase
• luminol oxidation
• reversed micelles
• Aerosol OT
• Cortisol conjugate