An enzyme-level benchmark based on environmental bacterial laccases for predicting contaminant fate in water

Abstract Bacterial laccases are widespread multicopper oxidases whose roles in the fate of anthropogenic chemicals in aquatic environments remain poorly understood. Here, we integrate metagenomic analysis, a miniaturized high-throughput assay and machine learning to establish an enzyme-level benchmark for predicting biotransformation of wastewater-relevant trace organic contaminants by laccase-mediator systems. Using a laccase from an ammonia-oxidizing bacterium as a model enzyme, we screened 183 compounds and identified 38 that underwent significant removal. Following phylogenetic analysis of environmental homologs, we expressed and purified two additional laccases from the bacterial methanotrophic phylum Methylmirabilota and an archaeal phylum Thermoproteota , demonstrating the activity of this enzyme family across domains. Graph convolutional network models trained on the dataset achieved up to 78% accuracy in classifying degradable versus persistent chemicals, while quantum-chemical descriptors highlighted key electronic properties governing oxidation. This bottom-up approach to enzyme-chemical interactions establishes a trajectory towards predicting contaminant persistence in engineered and natural waters.