Exploiting the potential of metal and solvent tolerant laccase from Tricholoma giganteum AGDR1 for the removal of pesticides

Laccase from previously reported hardwood degrading fungus, Tricholoma giganteum AGDR1, was isolated, identified at molecular level, biochemically characterized and also utilized for pesticide degradation. Laccase gene is comprised of 3752 bp, which encompassed 742-bp of 5′ flanking upstream sequence with 12 introns and 12 exons. Mature enzyme possesses 391 amino acids and signal peptide, which is determined to be monomeric protein with an apparent molecular weight of 41 kDa and 6.45 pI. Higher optimal activities were observed at 45 °C and pH 3.0 and surprisingly, it exhibited more than 20% of relative activity at pH 1.5. Purified laccase was tolerant to 100 mM of metals (i.e. Se, Pb, Cu, Cr and Cd), organic solvents (ethyl acetate, methanol, ethanol and acetone) and potent inhibitors (hydroxylamine, thiourea, NaF and Na-azide) as compared to reported laccases. It was able to degrade 29%, 7% and 72% of chlorpyrifos, profenofos and thiophanate methyl within 15 h, respectively. Molecular docking analysis revealed that higher binding efficacy of these pesticides is observed with H83, H320, A95, V384, and P366 which are presented near to the catalytic site. Based on the results, T. giganteum AGDR1 laccase can be applied for the potential remediation and industrial applications under harsh conditions.