Salt leverages polyethylene terephthalate hydrolase (PETase) enzymatic activity via the predicted conformational switch

Plastic pollution spawned a global challenge caused by the environmental accumulation of polyethylene terephthalate (PET) plastics. Ongoing remediation efforts using microbial and engineered PET hydrolyzing enzymes (PETases) are hindered by slow depolymerization activities. Here, we report the optimized reaction conditions that leveraged the PETase hydrolase activity 2 to 3.8-fold in the presence of high NaCl concentrations. Molecular dynamics simulations (MDS) were applied to model salt-dependent conformational changes of the PETase enzyme bound to a 3-unit PET polymer. MDS demonstrated that PETase interaction with flanking polymer units exhibited a striking structural disparity at low and high salt concentrations. At low salt concentrations, flanking polymer units displayed significant bending. In contrast, flanking units extended at high salt concentrations, thus residues D206, H237, and S160 of the catalytic triad positioned in close vicinity of the scissile ester bond of the polymer substrate. The resulting high salt-specific enzyme/substrate geometry can potentially facilitate hydrolysis. We theorized that a salt-dependent conformational switch could attenuate the enzyme to a broad range of natural and artificial polymers consumed as carbon sources. Altogether, new knowledge may advance the engineering of PET hydrolase enzymes and benefit bioconversion programs. ### Competing Interest Statement The authors have declared no competing interest. * Abbreviations : PET : polyethylene terephthalate PETase : PET hydrolase MD : molecular dynamics MHET : mono-2-hydroxyethyl acid BHET : bis-2-hydroxyethyl terephthalic acid p-NPA : P-nitrophenyl acetate HEMT : (2-hydroxyethyl) 4-methyl terephthalate