The method of integrated kinetics and its applicability to the exo-glycosidase-catalyzed hydrolysis of p-nitrophenyl glycosides

被引：4

作者：

Borisova, Anna S. ^{[1
,2
]}

Reddy, Sumitha K. ^{[3
]}

Ivanen, Dina R. ^{[1
]}

Bobrov, Kirill S. ^{[1
]}

Eneyskaya, Elena V. ^{[1
]}

Rychkov, Georgy N. ^{[1
,5
]}

Sandgren, Mats ^{[2
]}

Stalbrand, Henrik ^{[3
]}

Sinnott, Michael L. ^{[4
]}

Kulminskaya, Anna A. ^{[1
,5
]}

Shabalin, Konstantin A. ^{[1
,5
]}

机构：

[1] Natl Res Ctr Kurchatov Inst, BP Konstantinov Petersburg Nucl Phys Inst, Gatchina 188300, Orlova Roscha, Russia

[2] Swedish Univ Agr Sci, Dept Chem & Biotechnol, Uppsala, Sweden

[3] Lund Univ, Dept Biochem & Struct Biol, S-22100 Lund, Sweden

[4] Univ Huddersfield, Dept Chem Sci, Huddersfield HD1 3DH, W Yorkshire, England

[5] St Petersburg State Polytech Univ, St Petersburg 195251, Russia

来源：

CARBOHYDRATE RESEARCH | 2015年 / 412卷

基金：

俄罗斯基础研究基金会;

关键词：

Integrated kinetics; Retaining glycoside hydrolase; Mutarotation; ALPHA-GALACTOSIDASE; BETA-GALACTOSIDASE; LACTOSE HYDROLYSIS; CRYSTAL-STRUCTURE; RATE EQUATIONS; XYLOSIDASE; CONSTANTS;

D O I：

10.1016/j.carres.2015.03.021

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

In the present work we suggest an efficient method, using the whole time course of the reaction, whereby parameters k(cat), K-m and product K-I for the hydrolysis of a p-nitrophenyl glycoside by an exo-acting glycoside hydrolase can be estimated in a single experiment. Its applicability was demonstrated for three retaining exo-glycoside hydrolases, beta-xylosidase from Aspergillus awamori, beta-galactosidase from Penicillium sp. and alpha-galactosidase from Thermotoga maritima (TmGalA). During the analysis of the reaction course catalyzed by the TmGalA enzyme we had observed that a non-enzymatic process, mutarotation of the liberated alpha-D-galactose, affected the reaction significantly. (C) 2015 Elsevier Ltd. All rights reserved.

引用

页码：43 / 49

页数：7

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