Optimisation of a high-throughput fluorescamine assay for detection of N-acyl-l-homoserine lactone acylase activity

N-acyl-l-homoserine lactone (AHL) acylases are a well-known group of enzymes that disrupt quorum sensing in Gram-negative bacteria by degrading AHL signalling molecules. This degradation of signalling molecules (termed 'quorum quenching') has potential uses in the prevention or reduction of biofilm formation and/or bacterial infections. Therefore, there is a great deal of interest in the identification and characterisation of quorum quenching enzymes. Here, we present an optimised fluorescamine-based assay for the detection of AHL acylase activity and demonstrate it can be used in a high-throughput screening format. •N-acyl-l-homoserine lactone (AHL) acylases degrade quorum sensing molecules.•A fluorescamine-based assay for the detection of homoserine lactone was optimised.•This assay can be used to screen potential acylases for activity towards AHLs.