Evaluation of the Numbers of Functional Groups Introduced into Horseradish Peroxidase in Reactions with Four Heterobifunctional Reagents

Abstract

Numbers of functional groups introduced into horseradish peroxidase in reactions with four heterobifunctional reagents, S-acetylmercaptosuccinic anhydride, N-(ε-maleimidocaproyloxy)succinimide, N-succinimidyl 3-(2-pyridyldithio)propionate and N-(3-bromoacetamido-n-propionoyloxy)succinimide, were investigated. In the reactions at large molar ratios of the reagents to the enzyme in neutral and slightly alkaline media, maximum numbers of the incorporated functional groups were approximately 2.2 : two reactive amino groups are present in the enzyme molecule.