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Hydrocarbon and Lipid Microbiology Protocols pp 37–69Cite as

Screening for Enantioselective Lipases

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Abstract

Many lipolytic enzymes are enantioselective thus being able to distinguish between two enantiomers of a given racemic substrate. This property together with ample availability and comparatively easy handling makes lipolytic enzymes the most widely used class of biocatalysts in the chemical and pharmaceutical industries. However, lipase activity as well as selectivity is often negligible towards typical industrial substrates which usually do not resemble natural ones. Therefore, suitable enzymes must first be identified, usually by activity-based screening methods which, however, differ in reliability, throughput and surrogate function. Here, we describe important parameters determining the reliability and reproducibility of such screening systems for five different assays in detail. Moreover, comprehensive protocols for the synthesis of enantiopure lipase substrates and their use for screening of enantioselective lipases are provided.

Keywords:

  • Adrenaline assay
  • Agar plate assay
  • Colorimetric assay
  • Fluorometric assay
  • Quick E assay

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  • DOI: 10.1007/8623_2016_218
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Author information

Authors and Affiliations

  1. Institute of Bioorganic Chemistry, Heinrich-Heine-University Düsseldorf, Forschungszentrum Jülich, D-52426, Jülich, Germany

    Thomas Classen, Benjamin Lauinger & Jörg Pietruszka

  2. Institute of Bio- and Geosciences IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, D-52426, Jülich, Germany

    Thomas Classen, Jörg Pietruszka & Karl-Erich Jaeger

  3. Institute of Molecular Enzyme Technology, Heinrich-Heine-University Düsseldorf, Forschungszentrum Jülich, D-52426, Jülich, Germany

    Filip Kovacic & Karl-Erich Jaeger

Authors
  1. Thomas Classen
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  2. Filip Kovacic
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  3. Benjamin Lauinger
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  4. Jörg Pietruszka
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  5. Karl-Erich Jaeger
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Corresponding author

Correspondence to Karl-Erich Jaeger .

Editor information

Editors and Affiliations

  1. School of Biological Sciences, University of Essex Dept. Biological Sciences, Colchester, Essex, United Kingdom

    Terry J. McGenity

  2. Institute of Microbiology, Technical University Braunschweig, Braunschweig, Germany

    Kenneth N. Timmis

  3. Department of Biology, University of the Balearic Islands and Mediterranean Institute for Advanced Studies (IMEDEA, UIB-CSIC), Palma de Mallorca, Spain

    Balbina Nogales

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Classen, T., Kovacic, F., Lauinger, B., Pietruszka, J., Jaeger, KE. (2016). Screening for Enantioselective Lipases. In: McGenity, T., Timmis, K., Nogales, B. (eds) Hydrocarbon and Lipid Microbiology Protocols. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8623_2016_218

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  • DOI: https://doi.org/10.1007/8623_2016_218

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