Formation and nature of non-extractable residues of emerging organic contaminants in humic acids catalyzed by laccase

Formation of non-extractable residues (NERs) is the major fate of most environmental organic contaminants in soil, however, there is no direct evidence yet to support the assumed physical entrapment of NERs (i.e., type I NERs) inside soil humic substances. Here, we used 14C-radiotracer and silylation techniques to analyze NERs of six emerging and traditional organic contaminants formed in a suspension of humic acids (HA) under catalysis of the oxidative enzyme laccase. Laccase induced formation of both type I and covalently bound NERs (i.e., type II NERs) of bisphenol A, bisphenol F, and tetrabromobisphenol A to a large extent, and of bisphenol S (BPS) and sulfamethoxazole (SMX) to a less extent, while no induction for phenanthrene. The type I NERs were formed supposedly owing to laccase-induced alteration of primary (active groups) and secondary (conformation) structure of humic supramolecules, contributing surprisingly to large extents (23.5%-65.7%) to the total NERs, particularly for BPS and SMX, which both were otherwise not transformed by laccase catalysis. Electron-withdrawing sulfonyl group and bromine substitution significantly decreased amount and kinetics of NER formation, respectively. This study provides the first direct evidence for the formation of type I NERs in humic substances and implies a "Trojan horse" effect of such NERs in the environment.