7QE2
Crystal structure of D-glucuronic acid bound to SN243
Summary for 7QE2
| Entry DOI | 10.2210/pdb7qe2/pdb |
| Descriptor | SN243, beta-D-glucopyranuronic acid, ACETATE ION, ... (6 entities in total) |
| Functional Keywords | enzyme discovery, carbohydrate-active enzymes (cazy), protein engineering, functional metagenomics), hydrolase |
| Biological source | unidentified |
| Total number of polymer chains | 2 |
| Total formula weight | 166823.77 |
| Authors | Neun, S.,Brear, P.,Campbell, E.,Omari, K.,Wagner, O.,Hyvonen, M.,Hollfelder, F. (deposition date: 2021-12-01, release date: 2022-10-12, Last modification date: 2024-10-23) |
| Primary citation | Neun, S.,Brear, P.,Campbell, E.,Tryfona, T.,El Omari, K.,Wagner, A.,Dupree, P.,Hyvonen, M.,Hollfelder, F. Functional metagenomic screening identifies an unexpected beta-glucuronidase. Nat.Chem.Biol., 18:1096-1103, 2022 Cited by PubMed Abstract: The abundance of recorded protein sequence data stands in contrast to the small number of experimentally verified functional annotation. Here we screened a million-membered metagenomic library at ultrahigh throughput in microfluidic droplets for β-glucuronidase activity. We identified SN243, a genuine β-glucuronidase with little homology to previously studied enzymes of this type, as a glycoside hydrolase 3 family member. This glycoside hydrolase family contains only one recently added β-glucuronidase, showing that a functional metagenomic approach can shed light on assignments that are currently 'unpredictable' by bioinformatics. Kinetic analyses of SN243 characterized it as a promiscuous catalyst and structural analysis suggests regions of divergence from homologous glycoside hydrolase 3 members creating a wide-open active site. With a screening throughput of >10 library members per day, picolitre-volume microfluidic droplets enable functional assignments that complement current enzyme database dictionaries and provide bridgeheads for the annotation of unexplored sequence space. PubMed: 35799064DOI: 10.1038/s41589-022-01071-x PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.15 Å) |
Structure validation
Download full validation report
