Miniaturized whole-cell bacterial bioreporter assay for identification of quorum sensing interfering compounds

• Published in: Journal of microbiological methods 2018-11, Vol.154, p.40-45
• Main Authors: Ilina, Polina, Ma, Xiaochu, Sintim, Herman O., Tammela, Päivi
• Format: Article
• Language: English
• Subjects: Archaeal Proteins - genetics; Assay miniaturization; Autoinducer-2; Bacteria - drug effects; Bacteria - genetics; Bacteria - metabolism; beta-Galactosidase - metabolism; Biological Assay - methods; Bioreporter strain; Carrier Proteins - drug effects; Carrier Proteins - genetics; Carrier Proteins - metabolism; Drug Discovery - methods; Drug Resistance, Bacterial; Escherichia coli - drug effects; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli Proteins - drug effects; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; High throughput screening; Pentanones - antagonists & inhibitors; Quorum Sensing; Quorum sensing inhibitors; Signal Transduction
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2018.10.005

The continuing emergence and spread of antibiotic-resistant bacteria is worrisome and new strategies to curb bacterial infections are being sought. The interference of bacterial quorum sensing (QS) signaling has been suggested as a prospective antivirulence strategy. The AI-2 QS system is present in multiple bacterial species and has been shown to be correlated with pathogenicity. To facilitate the discovery of novel compounds interfering with AI-2 QS, we established a high-throughput setup of whole-cell bioreporter assay, which can be performed in either 96- or 384-well format. Agonistic or antagonistic activities of the test compounds against Escherichia coli LsrB-type AI-2 QS system are monitored by measuring the level of β-galactosidase expression. A control strain expressing β-galactosidase in quorum sensing-independent manner is included into the assay for false-positive detection. •Bacterial quorum sensing is a potential target in antivirulence drug discovery.•E. coli- based bioreporter assay to detect autoinducer-2 quorum sensing interference.•High-throughput 96- and 384-well formats•Efficiently detects antagonistic and agonistic activity of drug candidates.•Detects unspecific inhibitors (false-positives)