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High-Throughput Screening for Thermostable Polyester Hydrolases

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Metagenomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2555))

Abstract

Due to the promise of more sustainable recycling of plastics through biocatalytic degradation, the search for and engineering of polyester hydrolases have become a thriving field of research. Furthermore, among other methods, halo formation assays have become popular for the detection of polyester-hydrolase activity. However, established halo-formation assays are limited in their ability to screen for thermostable enzymes, which are particularly important for efficient plastic degradation. The incubation of screening plates at temperatures above 50 °C leads to cell lysis and death. Therefore, equivalent master plates are commonly required to maintain and identify the active strains found on the screening plates. This replica plating procedure necessitates 20- to 60-fold more plates than our method, assuming the screened library is transferred to 384-well microtiter plates or 96-well microtiter plates, respectively, to organize the colonies in a retraceable manner, thus significantly lowering throughput. Here, we describe a halo formation assay that is designed to screen thermostable polyesterases independent of master plates and colony replication, thereby markedly reducing the workload and increasing the throughput.

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Acknowledgements

We would like to thank Covestro and CSC JÄKLECHEMIE for providing us with Impranil® DLN W50 for our research.

Author information

Authors and Affiliations

  1. Department of Biotechnology & Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Greifswald, Germany

    Yannick Branson, Christoffel P. S. Badenhorst, Lara Pfaff, Carolin Buchmann, Ren Wei & Uwe T. Bornscheuer

Authors
  1. Yannick Branson
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  2. Christoffel P. S. Badenhorst
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  3. Lara Pfaff
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  4. Carolin Buchmann
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  5. Ren Wei
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  6. Uwe T. Bornscheuer
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Corresponding authors

Correspondence to Ren Wei or Uwe T. Bornscheuer .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Biotechnology, University of Hamburg, Hamburg, Germany

    Wolfgang R. Streit

  2. Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Göttingen, Germany

    Rolf Daniel

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Branson, Y., Badenhorst, C.P.S., Pfaff, L., Buchmann, C., Wei, R., Bornscheuer, U.T. (2023). High-Throughput Screening for Thermostable Polyester Hydrolases. In: Streit, W.R., Daniel, R. (eds) Metagenomics. Methods in Molecular Biology, vol 2555. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2795-2_11

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  • DOI: https://doi.org/10.1007/978-1-0716-2795-2_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2794-5

  • Online ISBN: 978-1-0716-2795-2

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