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Measuring Enzyme Kinetics of Glycoside Hydrolases Using the 3,5-Dinitrosalicylic Acid Assay

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1588)

Abstract

Use of the 3,5-dinitrosalicylic acid reagent allows the simple and rapid quantification of reducing sugars. The method can be used for analysis of biological samples or in the characterization of enzyme reactions. Presented here is an application of the method in measuring the kinetics of a glycoside hydrolase reaction, including the optimization of the DNSA reagent, and the production of a standard curve of absorbance and sugar concentration.

Key words

Reducing sugars Enzyme kinetics Glycoside hydrolase UV–visible spectrophotometry 3,5-dinitrosalicylic acid 

Notes

Acknowledgments

The author thanks the support of the Knut and Alice Wallenberg Foundation through the Wallenberg Wood Science Centre, Sweden, as well as the KTH Carbohydrate Materials Consortium (CarboMat) supported by the Swedish Research Council FORMAS.

References

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Division of Glycoscience, School of Biotechnology, KTH, Royal Institute of TechnologyAlbaNova University CentreStockholmSweden

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