Quorum Sensing pp 311-323 | Cite as

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

  • Maybelle Kho Go
  • Jeng Yeong Chow
  • Wen Shan Yew
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

N-Acyl-homoserine lactones N-Acyl-homoserine lactonases Directed evolution Bioluminescence 

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Maybelle Kho Go
    • 1
  • Jeng Yeong Chow
    • 1
  • Wen Shan Yew
    • 1
  1. 1.Synthetic Biology for Clinical and Technological Innovation (SynCTI), Centre for Life SciencesNational University of SingaporeSingaporeSingapore

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