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

Abstract

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.

Keywords

Integrated kinetics; Retaining glycoside hydrolase; Mutarotation

Published in

Carbohydrate Research
2015, volume: 412, pages: 43-49
Publisher: ELSEVIER SCI LTD

SLU Authors

• Borisova, Anna

  • National Research Centre - Kurchatov Institute
  • The Department of Chemistry and Biotechnology, Swedish University of Agricultural Sciences
    

• Sandgren, Mats

  • The Department of Chemistry and Biotechnology, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Organic Chemistry
Biophysics

Publication identifier

• DOI: https://doi.org/10.1016/j.carres.2015.03.021

Permanent link to this page (URI)

https://res.slu.se/id/publ/68450