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

Gregory Arnal; Mohamed A Attia; Jathavan Asohan; Zhenhuan Lei; Benedikt Golisch; Harry Brumer

doi:10.1007/978-1-0716-3151-5_1

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

Methods Mol Biol. 2023:2657:3-14. doi: 10.1007/978-1-0716-3151-5_1.

Authors

Gregory Arnal¹, Mohamed A Attia^{1

2}, Jathavan Asohan¹, Zhenhuan Lei^{1

2}, Benedikt Golisch^{1

2}, Harry Brumer^{3

4

5}

Affiliations

¹ Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.
² Department of Chemistry, University of British Columbia, Vancouver, BC, Canada.
³ Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada. brumer@msl.ubc.ca.
⁴ Department of Chemistry, University of British Columbia, Vancouver, BC, Canada. brumer@msl.ubc.ca.
⁵ Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada. brumer@msl.ubc.ca.

PMID: 37149519
DOI: 10.1007/978-1-0716-3151-5_1

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 that enables the rapid, parallel determination of GH kinetics in applications ranging from initial activity screening and assay optimization to precise Michaelis-Menten analysis.

Keywords: Carbohydrate-active enzymes (CAZymes); Copper-bicinchoninic acid (BCA); Enzymology; Glycosidase; Glycoside hydrolase (GH); Polysaccharide; Reducing sugar.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Copper
Glycoside Hydrolases*
Polysaccharides
Quinolines*

Substances

Glycoside Hydrolases
bicinchoninic acid
Copper
Quinolines
Polysaccharides