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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; Krishnaswamyreddy, Sumitha LU ; Ivanen, Dina R; Bobrov, Kirill S; Eneyskaya, Elena V; Rychkov, Georgy N; Sandgren, Mats; Stålbrand, Henrik LU ; Sinnott, Michael L and Kulminskaya, Anna A, et al. (2015) In Carbohydrate Research 412. p.43-49
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.
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publication status
published
subject
in
Carbohydrate Research
volume
412
pages
43 - 49
publisher
Elsevier
external identifiers
  • pmid:26005928
  • wos:000357542000007
  • scopus:84929649802
ISSN
1873-426X
DOI
10.1016/j.carres.2015.03.021
language
English
LU publication?
yes
id
141591d3-7ee7-4201-a9de-af35c94b54c9 (old id 5442446)
date added to LUP
2015-06-23 17:40:36
date last changed
2017-03-19 03:14:51
@article{141591d3-7ee7-4201-a9de-af35c94b54c9,
  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.},
  author       = {Borisova, Anna S and Krishnaswamyreddy, Sumitha and Ivanen, Dina R and Bobrov, Kirill S and Eneyskaya, Elena V and Rychkov, Georgy N and Sandgren, Mats and Stålbrand, Henrik and Sinnott, Michael L and Kulminskaya, Anna A and Shabalin, Konstantin A},
  issn         = {1873-426X},
  language     = {eng},
  pages        = {43--49},
  publisher    = {Elsevier},
  series       = {Carbohydrate Research},
  title        = {The method of integrated kinetics and its applicability to the exo-glycosidase-catalyzed hydrolyses of p-nitrophenyl glycosides.},
  url          = {http://dx.doi.org/10.1016/j.carres.2015.03.021},
  volume       = {412},
  year         = {2015},
}