The method of integrated kinetics and its applicability to the exo-glycosidase-catalyzed hydrolyses of p-nitrophenyl glycosides
In: 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, 2015, S. 43-49
academicJournal
Zugriff:
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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The method of integrated kinetics and its applicability to the exo-glycosidase-catalyzed hydrolyses of p-nitrophenyl glycosides
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Autor/in / Beteiligte Person: | 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. ; Shabalin, Konstantin A. |
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Zeitschrift: | 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, 2015, S. 43-49 |
Veröffentlichung: | Elsevier, 2015 |
Medientyp: | academicJournal |
DOI: | 10.1016/j.carres.2015.03.021 |
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