2WNE

Mutant Laminarinase 16A cyclizes laminariheptaose

2WNE の概要

• エントリーDOI: 10.2210/pdb2wne/pdb
• 関連するPDBエントリー: 2CL2 2W39 2W52 2WLQ
• 分子名称: PUTATIVE LAMINARINASE, beta-D-glucopyranose-(1-3)-beta-D-glucopyranose-(1-3)-beta-D-glucopyranose-(1-3)-beta-D-glucopyranose-(1-3)-beta-D-glucopyranose-(1-3)-beta-D-glucopyranose-(1-3)-alpha-D-glucopyranose, alpha-D-mannopyranose-(1-2)-alpha-D-mannopyranose-(1-6)-[alpha-D-mannopyranose-(1-3)]alpha-D-mannopyranose-(1-6)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, ... (4 entities in total)
• 機能のキーワード: laminarin, family 16, cyclical poysaccharides, glycosyl hydrolase, beta sandwich, basidiomycete, beta-glucanase, gh7, gh16, lam16a, beta-1\, 6-glucan, hydrolase
• 由来する生物種: PHANEROCHAETE CHRYSOSPORIUM (WHITE-ROT FUNGUS)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34268.79

構造登録者

Vasur, J.,Kawai, R.,Andersson, E.,Widmalm, G.,Jonsson, K.H.M.,Hansson, H.,Engstrom, A.,Einarsson, E.,Forsberg, Z.,Igarashi, K.,Sandgren, M.,Samejima, M.,Stahlberg, J. (登録日: 2009-07-09, 公開日: 2010-01-26, 最終更新日: 2024-10-23)

主引用文献

Vasur, J.,Kawai, R.,Jonsson, K.H.M.,Widmalm, G.,Engstrom, A.,Frank, M.,Andersson, E.,Hansson, H.,Forsberg, Z.,Igarashi, K.,Samejima, M.,Sandgren, M.,Stahlberg, J.
Synthesis of Cyclic Beta-Glucan Using Laminarinase 16A Glycosynthase Mutant from the Basidiomycete Phanerochaete Chrysosporium.
J.Am.Chem.Soc., 132:1724-, 2010

PubMed Abstract: Glycosynthases are precise molecular instruments for making specifically linked oligosaccharides. X-ray crystallography screening of ligands bound to the 1,3(4)-beta-D-glucanase nucleophile mutant E115S of Phanerochaete chrysosporium Laminarinase 16A (Lam16A) showed that laminariheptaose (L7) bound in an arch with the reducing and nonreducing ends occupying either side of the catalytic cleft of the enzyme. The X-ray structure of Lam16A E115S in complex with alpha-laminariheptaosyl fluoride (alphaL7F) revealed how alphaL7F could make a nucleophilic attack upon itself. Indeed, when Lam16A E115S was allowed to react with alphaL7F the major product was a cyclic beta-1,3-heptaglucan, as shown by mass spectrometry. NMR confirmed uniquely beta-1,3-linkages and no reducing end. Molecular dynamics simulations indicate that the cyclic laminariheptaose molecule is not completely planar and that torsion angles at the glycosidic linkages fluctuate between two energy minima. This is the first report of a glycosynthase that joins the reducing and nonreducing ends of a single oligosaccharide and the first reported synthesis of cyclic beta-glucan.

PubMed: 20078120
DOI: 10.1021/JA909129B

実験手法

X-RAY DIFFRACTION (2.124 Å)