In Vitro and Computational Response of Differential Catalysis by BAFC 633 Laccase in Interaction with 2,4-D and Chlorpyrifos.

Enzymes secreted by white rot fungi (WRF), such as laccase, offer a promising approach for the treatment of hazardous xenobiotic compounds. This study conducted a comprehensive analysis of the impact of the pesticides 2,4-dichlorophenoxyacetic acid (2,4-D) and chlorpyrifos on the laccase of BAFC 633 through in vitro and bioinformatics analyses. The fungal strain was shown to be tolerant to both pesticides, with notable morphological and ultrastructural alterations in the mycelium. Laccase activity and two isoenzymes (53 and 70 kDa) were detected in all initial treatments. The laccase was concentrated for subsequent catalytic evaluation in the presence of both pesticides, showing high stability at a pH of 3.6 and a temperature range of 50-60 °C. The gene, corresponding to this laccase, was modeled, and its structure revealed a defined catalytic pocket validated with a drug score of 0.61. Molecular docking estimated affinity energies of -5.06 and -9.41 Kcal mol for 2,4-D and chlorpyrifos, respectively. Molecular Mechanics Poisson-Boltzmann Surface Area (MM/PBSA) analysis through 250 ns of molecular dynamics revealed stronger hydrophobic interactions of laccase with chlorpyrifos and highlighted the importance of residue His460 in stabilizing both complexes. Understanding the impact of these agrochemicals on the catalytic function of laccase is crucial for developing future biotechnological strategies involving this enzyme.