Conformational Selection in Biocatalytic Plastic Degradation by PETase

Due to the steric effects imposed by bulky polymers, formation of catalytically competent enzyme and substrate conformations is critical in biodegradation plastics. In poly(ethylene terephthalate) (PET), backbone adopts different conformations, gauche trans, coexisting extents amorphous crystalline regions. However, which conformation susceptible extent conformational changes required for expedient catalysis remain poorly understood. To overcome this obstacle, we utilized molecular dynamics simulations, docking, engineering concert with high-resolution microscopy imaging solid-state nuclear magnetic resonance (NMR) demonstrate importance selection biocatalytic plastic hydrolysis. Our results how single-amino acid substitutions Ideonella sakaiensis PETase can alter its landscape, significantly affecting relative abundance productive ground-state structures ready bind discrete conformers. We experimentally show an binds provide a model key residues involved recognition trans supported silico simulations. be used create trans-selective variant, resulting higher activity when combined all-trans PET-derived oligomeric substrate, stemming from both increased accessibility preference. work cements matching hydrolysis, that also noncanonical PET conducive degradation. Understanding contribution degradation could facilitate generation designer enzymes performance.