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.
CITATION STYLE
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 Methods in Molecular Biology (Vol. 1673, pp. 311–323). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7309-5_24
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