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

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Protein-Carbohydrate Interactions

Part of the book series: Methods in Molecular Biology ((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.

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References

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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.

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Authors and Affiliations

  1. Division of Glycoscience, School of Biotechnology, KTH, Royal Institute of Technology, AlbaNova University Centre, 106 91, Stockholm, Sweden

    Lauren S. McKee

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  1. Lauren S. McKee
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Correspondence to Lauren S. McKee .

Editor information

Editors and Affiliations

  1. Agriculture and Agri-Food Canada, Lethbridge, Alberta, Canada

    D. Wade Abbott

  2. Biotechnology Institute, University of Groningen, Groningen, The Netherlands

    Alicia Lammerts van Bueren

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McKee, L.S. (2017). Measuring Enzyme Kinetics of Glycoside Hydrolases Using the 3,5-Dinitrosalicylic Acid Assay. In: Abbott, D., Lammerts van Bueren, A. (eds) Protein-Carbohydrate Interactions. Methods in Molecular Biology, vol 1588. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6899-2_3

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  • DOI: https://doi.org/10.1007/978-1-4939-6899-2_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6898-5

  • Online ISBN: 978-1-4939-6899-2

  • eBook Packages: Springer Protocols

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