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Cellobiohydrolase (CBH) Activity Assays

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Cellulases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1796))

Abstract

Cellulosic biomass is the most abundant biopolymer on the earth. It has great potential to quench the thirst of liquid energy by producing biofuels and thus help to mitigate human reliance on fossil fuels. Although several cellulase activity assay methods have been used to disintegrate the glycosidic bonds, the appropriate selection of substrates and synergistic involvement of multiple enzymes in hydrolytic activity is not yet fully understood. The proper quantification of hydrolytic enzymes and hydrolysates is challenging because of the heterogeneity of cellulose, changes in enzyme-substrate ratio and the presence of some inhibitory compounds like cellobiose and cellodextran. In the glycosyl hydrolase (GH) family, cellobiohydrolase (CBH) is expected to disrupt the crystalline cellulose and release the sugar molecules. Several methods have been proposed for CBH assay with slight modification in substrate and quantification of hydrolysates. However, the Avicel method is still considered as the most promising and efficient hydrolytic technique so far. The most commonly used CBH assays including Avicel and other recent methods for proper quantification are outlined in this chapter. Also a qualitative screening of CBH producing bacteria using carboxymethyl cellulose (CMC) agar plates is described.

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Authors and Affiliations

  1. Department of Biology, Lakehead University, Thunder Bay, ON, Canada

    Hem Kanta Sharma & Wensheng Qin

  2. Department of Chemical and Biochemical Engineering, Western University, London, ON, Canada

    Chunbao (Charles) Xu

Authors
  1. Hem Kanta Sharma
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  2. Wensheng Qin
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  3. Chunbao (Charles) Xu
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Corresponding author

Correspondence to Wensheng Qin .

Editor information

Editors and Affiliations

  1. Section of Sustainable Biotechnology, Department of Chemistry and Bioscience, Aalborg University, Copenhagen, Denmark

    Mette Lübeck

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Sharma, H.K., Qin, W., Xu, C.(. (2018). Cellobiohydrolase (CBH) Activity Assays. In: Lübeck, M. (eds) Cellulases. Methods in Molecular Biology, vol 1796. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7877-9_14

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  • DOI: https://doi.org/10.1007/978-1-4939-7877-9_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7876-2

  • Online ISBN: 978-1-4939-7877-9

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