9G5G

Glycoside Hydrolase Family 157 from Labilibaculum antarcticum (LaGH157) in complex with Laminaribiose

9G5G の概要

• エントリーDOI: 10.2210/pdb9g5g/pdb
• 関連するBIRD辞書のPRD_ID: PRD_900024
• 分子名称: Glycoside hydrolase family 2 catalytic domain-containing protein, beta-D-glucopyranose-(1-3)-beta-D-glucopyranose, MALONIC ACID, ... (7 entities in total)
• 機能のキーワード: glycoside hydrolase, endo-beta-1, 3-glucanase, cazyme, ligand complex, hydrolase
• 由来する生物種: Labilibaculum antarcticum
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 248079.94

構造登録者

Caseiro, C.,Alves, V.D.,Carvalho, A.L.,Bule, P. (登録日: 2024-07-16, 公開日: 2025-05-21)

主引用文献

Caseiro, C.,McGregor, N.G.S.,Alves, V.D.,Carvalho, A.L.,Romao, M.J.,Davies, G.J.,Fontes, C.M.G.A.,Bule, P.
Family GH157 enzyme exhibits broad linkage tolerance and a dual endo/exo-beta-glucanase activity on beta-glucans.
Int.J.Biol.Macromol., 282:137402-137402, 2024

PubMed Abstract: The structural and chemical diversity of β-glucans is reflected on the variety of essential biological roles tackled by these polysaccharides. This natural heterogeneity requires an elaborate assortment of enzymatic mechanisms to assemble, degrade or modify, as well as to extract their full biotechnological potential. Recent metagenomic efforts have provided an unprecedented growth in potential new biocatalysts, most of which remain unconfirmed or uncharacterized. Here we report the first biochemical and structural characterization of two bacterial β-glucanases from the recently created glycoside hydrolase family 157 (LaGH157 and BcGH157) and investigate their molecular basis for substrate hydrolysis. Structural analysis by X-ray crystallography revealed that GH157 enzymes belong to clan GH-A, possessing a (β/α)-barrel fold catalytic domain, two β-sandwich accessory domains and two conserved catalytic glutamates residues, with relative positions compatible with a retaining mechanism of hydrolysis. Specificity screening and enzyme kinetics suggest that the enzymes prefer mixed-linkage glucans over β-1,3-glucans. Activity screening showed that both enzymes exhibit pH optimum at 6.5 and temperature optimum for LaGH157 and BcGH157 at 25 °C and 48 °C, respectively. Product analysis with HPAEC-PAD and LC-MS revealed that both enzymes are endo-1,3(4)-β-glucanases, capable of cleaving β-1,3 and β-1,4-linked glucoses, when preceded by a β-1,3 linkage. Moreover, BcGH157 needs a minimum of 4 subsites occupied for hydrolysis to occur, while LaGH157 only requires 3 subsites. Additionally, LaGH157 possesses exohydrolytic activity on β-1,3 and branching β-1,6 linkages. This unusual bifunctional endo-1,3(4)/exo-1,3-1,6 activity constitutes an expansion on our understanding of β-glucan deconstruction, with the potential to inspire future applications.

PubMed: 39528173
DOI: 10.1016/j.ijbiomac.2024.137402

実験手法

X-RAY DIFFRACTION (2.71 Å)