Enzyme-catalyzed polymerization of 8-hydroxyquinoline-5-sulfonate byin situ nuclear magnetic resonance spectroscopy

Abstract

In this report, we describe the use of in situ NMR spectroscopy to elucidate the mechanism of horseradish peroxidase-catalyzed oxidative free-radical coupling of phenols. We demonstrate the potential of the technique for the polymerization of 8-hydroxyquinoline-5-sulfonate (HQS). Based on the structural changes, we establish the involvement of ortho- and para-position protons (to the hydroxyl group) in the oxidative free-radical coupling polymerization with their relative preferences. For example, in HQS, we establish that the positions 2, 4, and 7 are involved in the chemical bonding with the order of preference being 7 ≥ 2 > 4. Analyses of 13 C-NMR data suggest the formation of C-C- and C-O-C-type coupling bonds during enzymatic polymerization.