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Evaluation of Thermal Stability of Cellulosomal Hydrolases and Their Complex Formation

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Cellulases

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

Abstract

Enzymatic breakdown of plant biomass is an essential step for its utilization in biorefinery applications, and the products could serve as substrates for the sustainable and environmentally friendly production of fuels and chemicals. Toward this end, the incorporation of enzymes into polyenzymatic cellulosome complexes—able to specifically bind to and hydrolyze crystalline cellulosic materials, such as plant biomass—is known to increase the efficiency and the overall hydrolysis performance of a cellulase system. Despite their relative abundance in various mesophilic anaerobic cellulolytic bacteria, there are only a few reports of cellulosomes of thermophilic origin. However, since various biorefinery processes are favored by elevated temperatures, the development of thermophilic designer cellulosomes could be of great importance. Owing to the limited number of thermophilic cellulosomes, designer cellulosomes, composed of mixtures of mesophilic and thermophilic components, have been constructed. As a result, the overall thermal profile of the individual parts and the resulting complex has to be extensively evaluated. Here, we describe a practical guide for the determination of temperature stability for cellulases in the cellulosome complexes. The approach is also appropriate for other related enzymes, notably xylanases as well as other glycoside hydrolases. We provide detailed experimental procedures for the evaluation of the thermal stability of the individual designer cellulosome components and their complexes as well as protocols for the assessment of complex integrity at elevated temperatures.

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Author information

Authors and Affiliations

  1. Department of Biomolecular Sciences, The Weizmann Institute of Science, Rehovot, Israel

    Amaranta Kahn, Sarah Moraïs & Edward A. Bayer

  2. Microbiology Group, Department of Biology, National and Kapodistrian University of Athens, Athens, Greece

    Anastasia P. Galanopoulou & Dimitris G. Hatzinikolaou

Authors
  1. Amaranta Kahn
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  2. Anastasia P. Galanopoulou
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  3. Dimitris G. Hatzinikolaou
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  4. Sarah Moraïs
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  5. Edward A. Bayer
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Corresponding author

Correspondence to Edward A. Bayer .

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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Kahn, A., Galanopoulou, A.P., Hatzinikolaou, D.G., Moraïs, S., Bayer, E.A. (2018). Evaluation of Thermal Stability of Cellulosomal Hydrolases and Their Complex Formation. 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_12

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  • DOI: https://doi.org/10.1007/978-1-4939-7877-9_12

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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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