To build upon this recent discovery we conducted a broad exploration across naturally occurring sequence defined laccases for their ability to catalyze AFB1 degradation through a combination of genome mining in Brenda, Uniprot 33 , PHMMer , and review of previous literature . A total of approximately 89,000 genes were identified (Figure ) from which a set of 46 synthetic genes encoding the naturally occurring proteins were procured. The 46 genes covered had an average identity of 22% and a median identity of 14% to each other (Figure ). They covered the three largest clusters in the laccase family but were centered around previously hypothesized or identified enzymes in the family capable of oxidizing AFB1. All genes had signal peptides removed and were then synthesized, cloned into a pET-29b(+) vector (Novagen, USA), and transformed by heat-shock into Escherichia coli BLR(DE3) cells (Novagen, USA), which were grown overnight as 2mL seed cultures at 37°C in Terrific Broth. The cultures were expanded into 500 mL cultures and shaken overnight in Terrific Broth at 37°C. Once cells had reached optimal density they were induced with 1mM Isopropyl-β-D-thiogalactoside (IPTG) and 1mM CuSO4 at 20°C without shaking to limit the O2 concentration in the culture. Previous work has shown oxygen content of cultures can impact copper uptake into cells. These cultures were then lysed with sonication and purified using Ni-NTA Immobilized Metal Ion Chromatography before being eluted with phosphate buffered saline (PBS) containing 200 mM imidazole (pH 7.5). .