8S7Z

Urethanase umg-sp1 without inhibitor or substrate displays flexible active site loops

Summary for 8S7Z

• Entry DOI: 10.2210/pdb8s7z/pdb
• Descriptor: Urethanase umg-sp1 (2 entities in total)
• Functional Keywords: urethanase, amidase, amidase signature protein superfamily, catalytic triad, hydrolase
• Biological source: metagenome
• Total number of polymer chains: 4
• Total formula weight: 186105.28

Authors

Palm, G.J.,Graf, L.G.,Berndt, L.,Lammers, M. (deposition date: 2024-03-05, release date: 2024-07-24, Last modification date: 2024-10-09)

Primary citation

Bayer, T.,Palm, G.J.,Berndt, L.,Meinert, H.,Branson, Y.,Schmidt, L.,Cziegler, C.,Somvilla, I.,Zurr, C.,Graf, L.G.,Janke, U.,Badenhorst, C.P.S.,Konig, S.,Delcea, M.,Garscha, U.,Wei, R.,Lammers, M.,Bornscheuer, U.T.
Structural Elucidation of a Metagenomic Urethanase and Its Engineering Towards Enhanced Hydrolysis Profiles.
Angew.Chem.Int.Ed.Engl., 63:e202404492-e202404492, 2024

PubMed Abstract: While plastics like polyethylene terephthalate can already be degraded efficiently by the activity of hydrolases, other synthetic polymers like polyurethanes (PUs) and polyamides (PAs) largely resist biodegradation. In this study, we solved the first crystal structure of the metagenomic urethanase UMG-SP-1, identified highly flexible loop regions to comprise active site residues, and targeted a total of 20 potential hot spots by site-saturation mutagenesis. Engineering campaigns yielded variants with single mutations, exhibiting almost 3- and 8-fold improved activity against highly stable N-aryl urethane and amide bonds, respectively. Furthermore, we demonstrated the release of the corresponding monomers from a thermoplastic polyester-PU and a PA (nylon 6) by the activity of a single, metagenome-derived urethanase after short incubation times. Thereby, we expanded the hydrolysis profile of UMG-SP-1 beyond the reported low-molecular weight carbamates. Together, these findings promise advanced strategies for the bio-based degradation and recycling of plastic materials and waste, aiding efforts to establish a circular economy for synthetic polymers.

PubMed: 38948941
DOI: 10.1002/anie.202404492

Experimental method

X-RAY DIFFRACTION (2.67 Å)