Molecular insights into model polyester-based polyurethane impranil DLN degradation by TfCut2 cutinase
Abstract
The enzymatic degradation of synthetic polymers offers a sustainable route for plastic waste recycling, yet progress remains limited by an incomplete understanding of enzyme-polymer recognition and catalytic mechanisms. While cutinases have shown potential to degrade polyester-based polyurethanes (PURs), the molecular determinants of substrate binding and selectivity are still poorly resolved. In this study, we combined computational and experimental approaches to elucidate the interaction between Thermobifida fusca cutinase (TfCut2) and Impranil DLN, a polyester-based PUR widely used as a model substrate. NMR analysis refined the structural description of Impranil DLN, enabling the construction of accurate molecular models for simulations. Using computational analyses, we identified multiple catalytically competent TfCut2-Impranil complexes and char-acterised the key residues stabilising the substrate in the active site, including Y60, M131, and W155 forming an "aromatic clamp" near the oxyanion hole. The small differences in binding free energy across ligand poses suggest flexible substrate accommodation rather than a single preferred orientation, consistent with the enzyme's broad specificity. Guided by computational insights, targeted mutations near the active site were re-designed and experimentally tested. Notably, variants T61V, G62A, and T207D exhibited enhanced Impranil DLN degradation rates, with G62A showing over a twofold increase in initial rate and T207D demonstrating improved thermal stability and production yield. These findings underscore that efficient PUR degradation arises not from strong binding alone but from the enzyme's ability to guide the polymer into productive orientations while preserving catalytic geometry and turnover. This mechanistic understanding offers clearer design principles for improving PUR hydrolases.
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BibTeX
@article{raczynska2026molecular,
title = {{Molecular insights into model polyester-based polyurethane impranil DLN degradation by TfCut2 cutinase}},
author = {Agata Raczyńska and Oksana Kovalenko and Kinga Plasa and Tadeusz Bieg and Rajendra Sharma and Jeremy Esque and Katarzyna Papaj and Anna Kasprzycka and Artur Góra and Isabelle André},
journal = {International Journal of Biological Macromolecules},
year = {2026},
month = nov,
doi = {10.1016/j.ijbiomac.2026.151616},
url = {https://insa-toulouse.hal.science/hal-05850536v1},
note = {CC BY-ND 4.0},
}
RIS
TY - JOUR
TI - Molecular insights into model polyester-based polyurethane impranil DLN degradation by TfCut2 cutinase
AU - Agata Raczyńska
AU - Oksana Kovalenko
AU - Kinga Plasa
AU - Tadeusz Bieg
AU - Rajendra Sharma
AU - Jeremy Esque
AU - Katarzyna Papaj
AU - Anna Kasprzycka
AU - Artur Góra
AU - Isabelle André
PY - 2026
DA - 2026/11/03
JO - International Journal of Biological Macromolecules
DO - 10.1016/j.ijbiomac.2026.151616
UR - https://insa-toulouse.hal.science/hal-05850536v1
AB - The enzymatic degradation of synthetic polymers offers a sustainable route for plastic waste recycling, yet progress remains limited by an incomplete understanding of enzyme-polymer recognition and catalytic mechanisms. While cutinases have shown potential to degrade polyester-based polyurethanes (PURs), the molecular determinants of substrate binding and selectivity are still poorly resolved. In this study, we combined computational and experimental approaches to elucidate the interaction between Thermobifida fusca cutinase (TfCut2) and Impranil DLN, a polyester-based PUR widely used as a model substrate. NMR analysis refined the structural description of Impranil DLN, enabling the construction of accurate molecular models for simulations. Using computational analyses, we identified multiple catalytically competent TfCut2-Impranil complexes and char-acterised the key residues stabilising the substrate in the active site, including Y60, M131, and W155 forming an "aromatic clamp" near the oxyanion hole. The small differences in binding free energy across ligand poses suggest flexible substrate accommodation rather than a single preferred orientation, consistent with the enzyme's broad specificity. Guided by computational insights, targeted mutations near the active site were re-designed and experimentally tested. Notably, variants T61V, G62A, and T207D exhibited enhanced Impranil DLN degradation rates, with G62A showing over a twofold increase in initial rate and T207D demonstrating improved thermal stability and production yield. These findings underscore that efficient PUR degradation arises not from strong binding alone but from the enzyme's ability to guide the polymer into productive orientations while preserving catalytic geometry and turnover. This mechanistic understanding offers clearer design principles for improving PUR hydrolases.
SN - 0141-8130
ER -