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Development of a Sensitive Bioassay Method for Quorum Sensing Inhibitor Screening Using a Recombinant Agrobacterium tumefaciens  

Kim Yeon Hee (Department of Chemical and Biochemical Engineering and Institute for Environmental Technology and Industry, Pusan National University)
Kim Young Hee (Department of Chemical and Biochemical Engineering and Institute for Environmental Technology and Industry, Pusan National University)
Kim Jung Sun (Department of Biotechnology, Dongseo University)
Park Sunghoon (Department of Chemical and Biochemical Engineering and Institute for Environmental Technology and Industry, Pusan National University)
Publication Information
Biotechnology and Bioprocess Engineering:BBE / v.10, no.4, 2005 , pp. 322-328 More about this Journal
Abstract
Acylhomoserine lactones (AHLs) are known to be the triggering molecules in the quorum sensing mechanism of many gram-negative bacteria. In order to detect AHL inhibitors that are potential biofilm inhibitors, a convenient and sensitive bioassay was developed based on the $\beta$-galactosidase activity ($\beta$-GAL) of a recombinant Agrobacterium tumefaciens strain. A series of commercially available AHLs were tested for inducing $\beta$-GAL at varying concentrations in agar-plate and liquid cultures of the reporter strain. All AHLs tested exhibited a concentration­dependent induction, and octanoyl homoserine lactone (OHL) showed the highest sensitivity with a detection limit of 0.1 nM in the liquid culture assay. When fimbrolide, a known quorum sensing inhibitor, was added, induction of $\beta$-GAL by OHL was repressed. The repression at a constant OHL concentration was dependent on the fimbrolide concentration with the detection limit below 1 ppm, indicating that this assay is a sensitive method for screening AHL inhibitors.
Keywords
quorum sensing inhibitor; bioassay; acyl homoserine lactones; Agrobacterium tumefaciens; fimbrolide; biofilm inhibitor;
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