3SC7

First crystal structure of an endo-inulinase, from Aspergillus ficuum: structural analysis and comparison with other GH32 enzymes.

3SC7 の概要

• エントリーDOI: 10.2210/pdb3sc7/pdb
• 関連するPDBエントリー: 3RWK
• 分子名称: Inulinase, alpha-D-mannopyranose-(1-3)-alpha-D-mannopyranose-(1-6)-alpha-D-mannopyranose, beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (5 entities in total)
• 機能のキーワード: glycoside hydrolase family 32, endo-inulinase, glycosylation, cytosol, hydrolase
• 由来する生物種: Aspergillus ficuum
• 細胞内の位置: Secreted: O94220
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 57160.85

構造登録者

Housen, I.,Pouyez, J.,Roussel, G.,Mayard, A.,Vandamme, A.M.,Wouters, J.,Michaux, C. (登録日: 2011-06-07, 公開日: 2012-06-27, 最終更新日: 2024-11-06)

主引用文献

Pouyez, J.,Mayard, A.,Vandamme, A.M.,Roussel, G.,Perpete, E.A.,Wouters, J.,Housen, I.,Michaux, C.
First crystal structure of an endo-inulinase, INU2, from Aspergillus ficuum: Discovery of an extra-pocket in the catalytic domain responsible for its endo-activity.
Biochimie, 94:2423-2430, 2012

PubMed Abstract: Endo-inulinase is a member of glycosidase hydrolase family 32 (GH32) degrading fructans of the inulin type with an endo-cleavage mode and is an important class of industrial enzyme. In the present study, we report the first crystal structure of an endo-inulinase, INU2, from Aspergillus ficuum at 1.5 Å. It was solved by molecular replacement with the structure of exo-inulinase as search model. The 3D structure presents a bimodular arrangement common to other GH32 enzymes: a N-terminal 5-fold β-propeller catalytic domain with four β-sheets and a C-terminal β-sandwich domain organized in two β-sheets with five β-strands. The structural analysis and comparison with other GH32 enzymes reveal the presence of an extra pocket in the INU2 catalytic site, formed by two loops and the conserved motif W-M(I)-N-D(E)-P-N-G. This cavity would explain the endo-activity of the enzyme, the critical role of Trp40 and particularly the cleavage at the third unit of the inulin(-like) substrates. Crystal structure at 2.1 Å of INU2 complexed with fructosyl molecules, experimental digestion data and molecular modelling studies support these hypotheses.

PubMed: 22750808
DOI: 10.1016/j.biochi.2012.06.020

実験手法

X-RAY DIFFRACTION (1.5 Å)