Influence of natural organic matter on horseradish peroxidase-mediated removal of 17α-ethinylestradiol: Role of molecular weight

Ubiquitous natural organic matter (NOM) plays a crucial role in the peroxidase-mediated transformation of phenolic pollutants in aquatic environment. As a poorly defined polydispersed mixture of assorted organic substances with wide molecular weight (MW) distribution, NOM has far prevented researchers from finding out the primarily responsible components for the specific effect. In this work, MW fractionated NOMs (Mf-NOMs) were used to investigate their roles on horseradish peroxidase (HRP)-mediated transformation of 17α-ethinylestradiol (EE2). The removal rate of EE2 was restrained in the presence of pristine or Mf-NOMs, and the inhibitory mechanism was MW-dependent. Low Mf-NOMs restrained the enzymatic reaction by acting as competitive substrates, while high Mf-NOMs retained freely dissolved EE2 which reduced its availability for enzymatic reaction. The contribution of these two processes to the inhibition induced by pristine NOM was further quantified and found to be relevant to the reaction conditions, especially EE2 concentration. The findings of this work reveal more complex influences of NOM on the enzymatic reaction than ever demonstrated, which aids in understanding the fate of EE2 and other congener contaminants in natural and municipal water.