6HH9

Crystal structure of a two-domain esterase (CEX) active on acetylated mannans co-crystallized with mannopentaose

6HH9 の概要

• エントリーDOI: 10.2210/pdb6hh9/pdb
• 関連するPDBエントリー: 6HFZ
• 分子名称: GDSL-like protein, beta-D-mannopyranose-(1-4)-beta-D-mannopyranose-(1-4)-beta-D-mannopyranose-(1-4)-beta-D-mannopyranose, beta-D-mannopyranose-(1-4)-beta-D-mannopyranose-(1-4)-beta-D-mannopyranose, ... (4 entities in total)
• 機能のキーワード: esterase, carbohydrate, galactoglucomannan, deacetylation, biomass, gut flora, sugar binding protein
• 由来する生物種: Roseburia intestinalis L1-82
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 171784.97

構造登録者

Michalak, L.,La Rosa, S.L.,Rohr, A.K.,Aachmann, F.L.,Westereng, B. (登録日: 2018-08-27, 公開日: 2019-09-11, 最終更新日: 2024-01-17)

主引用文献

Michalak, L.,La Rosa, S.L.,Leivers, S.,Lindstad, L.J.,Rohr, A.K.,Lillelund Aachmann, F.,Westereng, B.
A pair of esterases from a commensal gut bacterium remove acetylations from all positions on complex beta-mannans.
Proc.Natl.Acad.Sci.USA, 117:7122-7130, 2020

PubMed Abstract: β-mannans and xylans are important components of the plant cell wall and they are acetylated to be protected from degradation by glycoside hydrolases. β-mannans are widely present in human and animal diets as fiber from leguminous plants and as thickeners and stabilizers in processed foods. There are many fully characterized acetylxylan esterases (AcXEs); however, the enzymes deacetylating mannans are less understood. Here we present two carbohydrate esterases, CE2 and CE17, from the Firmicute , which together deacetylate complex galactoglucomannan (GGM). The three-dimensional (3D) structure of CE17 with a mannopentaose in the active site shows that the CBM35 domain of CE17 forms a confined complex, where the axially oriented C2-hydroxyl of a mannose residue points toward the Ser41 of the catalytic triad. Cavities on the CE17 surface may accept galactosylations at the C6 positions of mannose adjacent to the mannose residue being deacetylated (subsite -1 and +1). In-depth characterization of the two enzymes using time-resolved NMR, high-performance liquid chromatography (HPLC), and mass spectrometry demonstrates that they work in a complementary manner. CE17 exclusively removes the axially oriented 2--acetylations on any mannose residue in an oligosaccharide, including double acetylated mannoses, while the CE2 is active on 3-, 4-, and 6-acetylations. Activity of CE2 is dependent on CE17 removing 2--acetylations from double acetylated mannose. Furthermore, transacetylation of oligosaccharides with the 2--specific CE17 provided insight into how temperature and pH affects acetyl migration on manno-oligosaccharides.

PubMed: 32170022
DOI: 10.1073/pnas.1915376117

実験手法

X-RAY DIFFRACTION (2.4 Å)