9KL9

crystal structure of a mutant Poly(Ethylene terephthalate) hydrolase

Summary for 9KL9

• Entry DOI: 10.2210/pdb9kl9/pdb
• Descriptor: Poly(Ethylene terephthalate) hydrolase (1 entity in total)
• Functional Keywords: hydrolase, mutant, dimer
• Biological source: bacterium HR29
• Total number of polymer chains: 2
• Total formula weight: 58635.54

Authors

Wang, H.,Feng, Y.,Du, X.Y. (deposition date: 2024-11-14, release date: 2025-10-01)

Primary citation

Wang, H.,Cun, Y.,Wang, M.,Du, X.,Yang, Z.,Wang, H.,Zhang, J.,Wang, P.,Feng, Y.,Zhu, Y.
Computational loop reconstruction based design of efficient PET hydrolases.
Commun Biol, 8:934-934, 2025

PubMed Abstract: Enzymatic PET depolymerization represents a promising approach for establishing a circular economy for PET plastics. Nonetheless, limitations in enzyme activity persist as significant challenges to its industrial application. In this research, the backbone structure of the β6-β7 loop for PET hydrolase Bhr-PETase derived from the thermophilic bacterium HR29 was reconstructed by introducing double mutations (H218N/F222M), resulting in variant Bhr-NMT with high thermal stability (T = 92.9 °C) and 87% increase in activity. Moreover, the loop reconstruction mutations are transplanted into the engineered PET hydrolases LCC-ICCG and Kubu-P, resulting variants LCC-ICCG-NM (T = 92.4 °C) and Kubu-P-NM (T = 92.9 °C). Under high substrate concentration (165 g kg) and an enzyme loading of 0.5 mg g, the designed variants Bhr-NMT, LCC-ICCG-NM, and Kubu-P-NM achieve an overall conversion of 93%, 90%, and 94%, respectively, outperforming the benchmark LCC-ICCG (85%). Notably, under reduced enzyme loading (0.3 mg g), Kubu-P-NM still reaches an overall conversion of 91%, which is significantly superior to benchmarks Kubu-P (83%) and LCC-ICCG (71%). Overall, the engineered PET hydrolases demonstrate significant potential for industrial PET waste recycling.

PubMed: 40527955
DOI: 10.1038/s42003-025-08364-6

Experimental method

X-RAY DIFFRACTION (2.9 Å)