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(Hyper)thermophilic Enzymes: Production and Purification

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Protein Downstream Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1129))

Abstract

The discovery of thermophilic and hyperthermophilic microorganisms, thriving at environmental temperatures near or above 100 °C, has revolutionized our ideas about the upper temperature limit at which life can exist. The characterization of (hyper)thermostable proteins has broadened our understanding and presented new opportunities for solving one of the most challenging problems in biophysics: how is structural stability and biological function maintained at high temperatures where “normal” proteins undergo dramatic structural changes? In our laboratory we have purified and studied many thermostable and hyperthermostable proteins in an attempt to determine the molecular basis of heat stability. Here, we present methods to express such proteins and enzymes in E. coli and provide a general protocol for overproduction and purification. The ability to produce enzymes that retain their stability and activity at elevated temperatures creates exciting opportunities for a wide range of biocatalytic applications.

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Author information

Authors and Affiliations

  1. Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands

    Pierpaolo Falcicchio, Mark Levisson, Servé W. M. Kengen & John van der Oost

  2. Center for Biomedical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA

    Sotirios Koutsopoulos

Authors
  1. Pierpaolo Falcicchio
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  2. Mark Levisson
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  3. Servé W. M. Kengen
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  4. Sotirios Koutsopoulos
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  5. John van der Oost
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Corresponding author

Correspondence to Sotirios Koutsopoulos .

Editor information

Editors and Affiliations

  1. Laboratory of Enzyme Technology, Agricultural University of Athens Department of Agr. Biotechnology, Athens, Greece

    Nikolaos E. Labrou

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Falcicchio, P., Levisson, M., Kengen, S.W.M., Koutsopoulos, S., van der Oost, J. (2014). (Hyper)thermophilic Enzymes: Production and Purification. In: Labrou, N. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 1129. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-977-2_34

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  • DOI: https://doi.org/10.1007/978-1-62703-977-2_34

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-976-5

  • Online ISBN: 978-1-62703-977-2

  • eBook Packages: Springer Protocols

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