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Directed Evolution of Quorum-Quenching Enzymes: A Method for the Construction of a Directed Evolution Platform and Characterization of a Quorum-Quenching Lactonase from Geobacillus kaustophilus

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Book cover Quorum Sensing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1673))

Abstract

A thermostable quorum-quenching lactonase from Geobacillus kaustophilus (GKL) was used as a template for in vitro directed evolution experiments. Here we describe the overexpression and purification of wild-type GKL, the construction of a quorum-quenching directed evolution platform using bioluminescence as a reporter, and the in vitro kinetic assay for the determination of kinetic parameters of wild-type GKL and its mutants.

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Author information

Authors and Affiliations

  1. Synthetic Biology for Clinical and Technological Innovation (SynCTI), Centre for Life Sciences, National University of Singapore, Level 2, 28 Medical Drive, Singapore, 117456, Singapore

    Maybelle Kho Go, Jeng Yeong Chow & Wen Shan Yew

Authors
  1. Maybelle Kho Go
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  2. Jeng Yeong Chow
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  3. Wen Shan Yew
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Corresponding author

Correspondence to Wen Shan Yew .

Editor information

Editors and Affiliations

  1. Department of Science, University Roma Tre, Rome, Italy

    Livia Leoni

  2. Department of Science, University Roma Tre, Rome, Italy

    Giordano Rampioni

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Go, M.K., Chow, J.Y., Yew, W.S. (2018). Directed Evolution of Quorum-Quenching Enzymes: A Method for the Construction of a Directed Evolution Platform and Characterization of a Quorum-Quenching Lactonase from Geobacillus kaustophilus . In: Leoni, L., Rampioni, G. (eds) Quorum Sensing. Methods in Molecular Biology, vol 1673. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7309-5_24

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  • DOI: https://doi.org/10.1007/978-1-4939-7309-5_24

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7308-8

  • Online ISBN: 978-1-4939-7309-5

  • eBook Packages: Springer Protocols

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