9EH6
Crystal Structure of AroC
Summary for 9EH6
| Entry DOI | 10.2210/pdb9eh6/pdb |
| Descriptor | Alpha/beta hydrolase fold-5 domain-containing protein (2 entities in total) |
| Functional Keywords | enzyme catalysis, pet, ssns, thermostability, hydrolase |
| Biological source | Caldilinea aerophila |
| Total number of polymer chains | 2 |
| Total formula weight | 49583.56 |
| Authors | Hu, Z.,Klupt, K.,Zechel, D.,Jia, Z.,Howe, G. (deposition date: 2024-11-22, release date: 2025-02-26, Last modification date: 2025-04-16) |
| Primary citation | Hu, Z.,Klupt, K.,Zechel, D.L.,Jia, Z.,Howe, G. Mining Thermophile Genomes for New PETases with Exceptional Thermostabilities Using Sequence Similarity Networks. Chembiochem, :e2500065-e2500065, 2025 Cited by PubMed Abstract: Enzymatic hydrolysis of polyethylene terephthalate (PET) is a promising technology for advancing a circular PET economy. Several PET-degrading α/β hydrolases have been identified, but the full potential of this enzyme family to catalyze PET hydrolysis remains largely unexplored. To address this, sequence similarity networks were employed to investigate the α/β hydrolase fold-5 subfamily (IPR029059) for new PETases. Priority was given to sequences from thermophiles, as thermostable enzymes are likely more suitable for industrial applications. Ten enzymes with ~20% sequence identity to the well-known LCC-PETase were identified, and seven were successfully overexpressed and purified for in vitro characterization. Each enzyme catalyzed the hydrolysis of p-nitrophenyl butyrate, a mimic of trimeric PET, and emulsified PET nanoparticles. Notably, three enzymes were also capable of hydrolyzing PET films. Novel PETases exhibited melting temperatures (Tm) exceeding 55 °C and only modest losses of activity after incubation at 70 °C for 24 hours. The crystal structure of AroC (Tm = 85 °C) was resolved to 2.2 Å, revealing several salt bridges that likely confer thermostability, and a unique loop that is conserved amongst the PETases described here. These novel enzymes will enable engineering campaigns to generate thermostable and catalytically efficient PETases for use as industrial biocatalysts. PubMed: 40069109DOI: 10.1002/cbic.202500065 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.25 Å) |
Structure validation
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