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

Borisova, Anna S., Reddy, Sumitha K., Ivanen, Dina R., Bobrov, Kirill S., Eneyskaya, Elena V., Rychkov, Georgy N., Sandgren, Mats, Stålbrand, Henrik, Sinnott, Michael L., Kulminskaya, Anna A. and Shabalin, Konstantin A. (2015) The method of integrated kinetics and its applicability to the exo-glycosidase-catalyzed hydrolyses of p-nitrophenyl glycosides. Carbohydrate Research, 412. pp. 43-49. ISSN 00086215

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Abstract

In the present work we suggest an efficient method, using the whole time course of the reaction, whereby parameters kcat, Km and product KI 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, β-xylosidase from Aspergillus awamori, β-galactosidase from Penicillium sp. and α-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 α-d-galactose, affected the reaction significantly.

Item Type: Article
Subjects: Q Science > Q Science (General)
Q Science > QD Chemistry
Schools: School of Applied Sciences
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Depositing User: Sara Taylor
Date Deposited: 25 Jun 2015 08:36
Last Modified: 25 Jun 2015 08:36
URI: http://eprints.hud.ac.uk/id/eprint/24876

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