Structure‐Guided Engineering of a Versatile Urethanase Improves Its Polyurethane Depolymerization Activity
Polyurethane (PUR), the fifth most prevalent synthetic polymer, substantially contributes to the global plastic waste problem. Biotechnology-based recycling methods have recently emerged as innovative solutions to plastic waste disposal and sparked interest among scientific communities and industrial stakeholders in discovering and designing highly active plastic-degrading enzymes. Here, the ligand-free crystal structure of UMG-SP2, a metagenome-derived urethanase with depolymerization activities, at 2.59 Å resolution, as well as its (co-)structures bound to a suicide hydrolase inhibitor and a short-chain carbamate substrate at 2.16 and 2.40 Å resolutions, respectively, is reported. Structural analysis and molecular dynamics simulations reveal that the flexible loop L3 consisting of residues 219-226 is crucial for regulating the hydrolytic activity of UMG-SP2. The semi-rational redesign of UMG-SP2 reveals superior variants, A141G and Q399A, exhibiting over 30.7- and 7.4-fold increased activities on polyester-PUR and a methylene diamine derivative of PUR, respectively, compared to the wild-type enzyme. These findings advance the understanding of the structure-function relationship of PUR-hydrolyzing enzymes, which hold great promise for developing effective industrial PUR recycling processes and mitigating the environmental footprint of plastic waste.
Authors
- Da’san M. M. Jaradat (ORCID: https://orcid.org/0000-0002-9853-3272)
- F. Lennartz (ORCID: https://orcid.org/0000-0001-5617-5502)
- Pedro Paiva (ORCID: https://orcid.org/0000-0002-7003-6055)
- Yannick Branson (ORCID: https://orcid.org/0000-0002-0282-7090)
- Pushpendra Singh (ORCID: https://orcid.org/0000-0002-7277-115X)
- Zhishuai Li (ORCID: https://orcid.org/0000-0003-3408-6300)
- Uwe T. Bornscheuer (ORCID: https://orcid.org/0000-0003-0685-2696)
- Lin Cong (ORCID: https://orcid.org/0000-0002-3263-8426)
- Maria J. Ramos (ORCID: https://orcid.org/0000-0002-7554-8324)
- Gert Weber (ORCID: https://orcid.org/0000-0003-3624-1060)
- Ren Wei (ORCID: https://orcid.org/0000-0003-3876-1350)
- Ulrike Garscha (ORCID: https://orcid.org/0000-0002-3645-7833)
- Wenshuo Li
- Pedro Alexandrino Fernandes
- You Song (ORCID: https://orcid.org/0000-0002-0289-7830)
- Thomas Bayer (ORCID: https://orcid.org/0000-0002-0656-3280)
- Weidong Liu (ORCID: https://orcid.org/0000-0001-7954-7700)
- Yangyang Chen (ORCID: https://orcid.org/0000-0001-9349-4246)
- Xu Han (ORCID: https://orcid.org/0000-0001-8319-8830)
- Louis Schmidt (ORCID: https://orcid.org/0000-0001-6977-9102)
Institutions
- Shenyang Pharmaceutical University (CN)
- Universidade do Porto (PT)
- Helmholtz-Zentrum Berlin für Materialien und Energie (DE)
- Al-Balqa Applied University (JO)
- Universität Greifswald (DE)
- Rede de Química e Tecnologia (PT)
- Tianjin Institute of Industrial Biotechnology (CN)
- University of Chinese Academy of Sciences (CN)
Publication Details
- Journal
- Advanced Science
- Published
- 2025-02-07
- DOI
- https://doi.org/10.1002/advs.202416019
- Citations
- 26
- Primary Topic
- biodegradable polymer synthesis and properties
- Type
- article
- Field-Weighted Citation Impact
- 15.53
Funders
- Salt Science Research Foundation
- European Commission
- National Natural Science Foundation of China
- Novo Nordisk
- Ministério da Ciência, Tecnologia e Ensino Superior
- Humboldt-Universität zu Berlin
- Freie Universität Berlin
- Nanjing Tech University
- Novo Nordisk Fonden
- Horizon 2020 Framework Programme
- Helmholtz-Zentrum Berlin für Materialien und Energie
- Leibniz-Gemeinschaft
- Fundação para a Ciência e a Tecnologia
- National Key Research and Development Program of China