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A Low-Volume, Parallel Copper-Bicinchoninic Acid (BCA) Assay for Glycoside Hydrolases

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Protein-Carbohydrate Interactions

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

Abstract

The quantitation of liberated reducing sugars by the copper-bicinchoninic acid (BCA) assay provides a highly sensitive method for the measurement of glycoside hydrolase (GH) activity, particularly on soluble polysaccharide substrates. Here, we describe a straightforward method adapted to low-volume polymerase chain reaction (PCR) tubes which enables the rapid, parallel determination of GH kinetics in applications ranging from initial activity screening and assay optimization, to precise Michaelis–Menten analysis.

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Acknowledgments

Funding from the Natural Sciences and Engineering Research Council of Canada (NSERC) via the Strategic Partnership Grants for Networks (for the Industrial Biocatalysis Network) and Discovery Grant programs is gratefully acknowledged. Equipment infrastructure was funded by the Canada Foundation for Innovation and the British Columbia Knowledge Development Fund. We thank Sean McDonald (Brumer group, UBC) for comments on an early version of this chapter.

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Authors and Affiliations

  1. Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, BC, V6T 1Z4, Canada

    Gregory Arnal, Mohamed A. Attia, Jathavan Asohan & Harry Brumer

  2. Department of Chemistry, University of British Columbia, 2185 East Mall, Vancouver, BC, Canada, V6T 1Z4

    Mohamed A. Attia & Harry Brumer

  3. Department of Biochemistry and Molecular Biology, University of British Columbia, 2185 East Mall, Vancouver, BC, Canada, V6T 1Z4

    Harry Brumer

Authors
  1. Gregory Arnal
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  2. Mohamed A. Attia
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  3. Jathavan Asohan
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  4. Harry Brumer
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Corresponding author

Correspondence to Harry Brumer .

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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Arnal, G., Attia, M.A., Asohan, J., Brumer, H. (2017). A Low-Volume, Parallel Copper-Bicinchoninic Acid (BCA) Assay for Glycoside Hydrolases. 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_1

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  • DOI: https://doi.org/10.1007/978-1-4939-6899-2_1

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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

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