1OJK

Anatomy of glycosynthesis: Structure and kinetics of the Humicola insolens Cel7BE197A and E197S glycosynthase mutants

1OJK の概要

• エントリーDOI: 10.2210/pdb1ojk/pdb
• 関連するPDBエントリー: 1A39 1DYM 1OJI 1OJJ 2A39
• 関連するBIRD辞書のPRD_ID: PRD_900005 PRD_900023
• 分子名称: ENDOGLUCANASE I, beta-D-glucopyranose-(1-4)-alpha-D-glucopyranose, beta-D-glucopyranose-(1-4)-beta-D-glucopyranose, ... (6 entities in total)
• 機能のキーワード: hydrolase, cellulose degradation, glycosynthase
• 由来する生物種: HUMICOLA INSOLENS
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 91132.01

構造登録者

Ducros, V.M.-A.,Tarling, C.A.,Zechel, D.L.,Brzozowski, A.M.,Frandsen, T.P.,Von Ossowski, I.,Schulein, M.,Withers, S.G.,Davies, G.J. (登録日: 2003-07-10, 公開日: 2004-01-07, 最終更新日: 2023-12-13)

主引用文献

Ducros, V.M.-A.,Tarling, C.A.,Zechel, D.L.,Brzozowski, A.M.,Frandsen, T.P.,Von Ossowski, I.,Schulein, M.,Withers, S.G.,Davies, G.J.
Anatomy of Glycosynthesis: Structure and Kinetics of the Humicola Insolens Cel7B E197A and E197S Glycosynthase Mutants
Chem.Biol., 10:619-, 2003

PubMed Abstract: The formation of glycoconjugates and oligosaccharides remains one of the most challenging chemical syntheses. Chemo-enzymatic routes using retaining glycosidases have been successfully harnessed but require tight kinetic or thermodynamic control. "Glycosynthases," specifically engineered glycosidases that catalyze the formation of glycosidic bonds from glycosyl donor and acceptor alcohol, are an emerging range of synthetic tools in which catalytic nucleophile mutants are harnessed together with glycosyl fluoride donors to generate powerful and versatile catalysts. Here we present the structural and kinetic dissection of the Humicola insolens Cel7B glycosynthases in which the nucleophile of the wild-type enzyme is mutated to alanine and serine (E197A and E197S). 3-D structures reveal the acceptor and donor subsites and the basis for substrate inhibition. Kinetic analysis shows that the E197S mutant is considerably more active than the corresponding alanine mutant due to a 40-fold increase in k(cat).

PubMed: 12890535
DOI: 10.1016/S1074-5521(03)00143-1

実験手法

X-RAY DIFFRACTION (1.5 Å)