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

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Author

  • Anna S Borisova
  • Sumitha Krishnaswamyreddy
  • Dina R Ivanen
  • Kirill S Bobrov
  • Elena V Eneyskaya
  • Georgy N Rychkov
  • Mats Sandgren
  • Henrik Stålbrand
  • Michael L Sinnott
  • Anna A Kulminskaya
  • Konstantin A Shabalin
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Summary, in English

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.

Publishing year

2015

Language

English

Pages

43-49

Publication/Series

Carbohydrate Research

Volume

412

Document type

Journal article

Publisher

Elsevier

Topic

  • Biological Sciences

Status

Published

Project

  • Galactomannan degradation by fungi and gut bacteria

ISBN/ISSN/Other

  • ISSN: 1873-426X