biostudies-literatureBiundo AStandortagentur TirolÖsterreichische ForschungsförderungsgesellschaftKungliga Tekniska HögskolanSvenska Forskningsrådet FormasVetenskapsrådetSteirische WirtschaftsförderungsgesellschaftBundesministerium für Wissenschaft, Forschung und WirtschaftVINNOVA36217-36226https://www.ebi.ac.uk/biostudies/studies/S-EPMC9074940biostudies-literatureUnknown9(62)The recalcitrance of plastics like nylon and other polyamides contributes to environmental problems (<i>e.g.</i> microplastics in oceans) and restricts possibilities for recycling. The fact that hitherto discovered amidases (EC 3.5.1. and 3.5.2.) only show no, or low, activity on polyamides currently obstructs biotechnological-assisted depolymerization of man-made materials. In this work, we capitalized on enzyme engineering to enhance the promiscuous amidase activity of polyesterases. Through enzyme design we created a reallocated water network adapted for hydrogen bond formation to synthetic amide backbones for enhanced transition state stabilization in the polyester-hydrolyzing biocatalysts <i>Humicola insolens</i> cutinase and <i>Thermobifida cellulosilytica</i> cutinase 1. This novel RSC advancesSwitched reaction specificity in polyesterases towards amide bond hydrolysis by enzyme engineering.PMC90749402017-0020102016-061602017-01116Syren POGuebitz GMRibitsch DBiundo ASubagia RMaurer MfalseSwitched reaction specificity in polyesterases towards amide bond hydrolysis by enzyme engineering.The recalcitrance of plastics like nylon and other polyamides contributes to environmental problems (<i>e.g.</i> microplastics in oceans) and restricts possibilities for recycling. The fact that hitherto discovered amidases (EC 3.5.1. and 3.5.2.) only show no, or low, activity on polyamides currently obstructs biotechnological-assisted depolymerization of man-made materials. In this work, we capitalized on enzyme engineering to enhance the promiscuous amidase activity of polyesterases. Through enzyme design we created a reallocated water network adapted for hydrogen bond formation to synthetic amide backbones for enhanced transition state stabilization in the polyester-hydrolyzing biocatalysts <i>Humicola insolens</i> cutinase and <i>Thermobifida cellulosilytica</i> cutinase 1. This novel 2019-01-01T00:00:00Z2019 Nov2026-05-31T21:58:05.569Z2025-02-19T03:24:13.983ZS-EPMC90749403554057510.1039/c9ra07519d