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Heterologous Overexpression, Purification, and In Vitro Characterization of AHL Lactonases

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Quorum Sensing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 692))

Abstract

Quorum-quenching enzymes are useful as biochemical tools and possible therapeutic proteins. One of the best-characterized families of these catalysts is the N-acyl-l-homoserine lactone (AHL) lactonases, which rely on a dinuclear metal ion active site to hydrolytically cleave the autoinducer’s lactone bond and inactivate signaling. A detailed understanding of how this enzyme works can help in the design of more selective and efficient reagents. To facilitate these studies, we describe a methodology to heterologously express, purify, and conduct in vitro characterization of several metalloforms of the AHL lactonase from Bacillus thuringiensis (AiiA). These procedures should be applicable to similar enzymes and will facilitate the production of more useful quorum-quenching reagents for biochemical studies and possible therapeutic applications.

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Acknowledgments

This work was supported in part by the Texas Advanced Research Program (Grant 003658-0018-2006) and the Robert A. Welch Foundation (Grant F-1572).

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Authors and Affiliations

  1. Division of Medicinal Chemistry, College of Pharmacy, University of Texas, Austin, TX, USA

    Pei W. Thomas & Walter Fast

Authors
  1. Pei W. Thomas
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  2. Walter Fast
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Editors and Affiliations

  1. Health Sciences Center, Department of Surgery, Texas Tech University, 4th Street 3601, Lubbock, 79430, Texas, USA

    Kendra P. Rumbaugh

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Thomas, P.W., Fast, W. (2011). Heterologous Overexpression, Purification, and In Vitro Characterization of AHL Lactonases. In: Rumbaugh, K. (eds) Quorum Sensing. Methods in Molecular Biology, vol 692. Humana Press. https://doi.org/10.1007/978-1-60761-971-0_20

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  • DOI: https://doi.org/10.1007/978-1-60761-971-0_20

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-970-3

  • Online ISBN: 978-1-60761-971-0

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