7QEE

SN243 mutant D415N bound to para-nitrophenyl-Beta-D-glucuronide

7QEE の概要

• エントリーDOI: 10.2210/pdb7qee/pdb
• 分子名称: SN243, ZINC ION, SULFATE ION, ... (6 entities in total)
• 機能のキーワード: enzyme discovery, carbohydrate-active enzymes (cazy), protein engineering, functional metagenomics), hydrolase
• 由来する生物種: Synthetic construct
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 166618.12

構造登録者

Neun, S.,Brear, P.,Campbell, E.,Omari, K.,Wagner, O.,Hyvonen, M.,Hollfelder, F. (登録日: 2021-12-02, 公開日: 2022-11-16, 最終更新日: 2024-11-20)

主引用文献

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

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: 35799064
DOI: 10.1038/s41589-022-01071-x

実験手法

X-RAY DIFFRACTION (2.374 Å)