Bioluminescence-based bioassays for rapid detection of nisin in food

Abstract

We have developed a method for determining ultralow amounts of nisin in food samples that is based on luminescent biosensor bacteria. Modified bacterial luciferase operon luxABCDE was placed under control of the nisin-inducible nisA promoter in plasmid pNZ8048, and the construct was transformed into Lactococcus lactis strains NZ9800 and NZ9000. The nisRK genes of these strains allow them to sense nisin and relay the signal to initiate transcription from nisA promoter. The resulting luminescence can be directly measured from living bacteria without the addition of exogenous substrates. Induction leads to detectable luminescence within ten minutes. Lyophilization of the biosensor cells produced viable and inducible sensor elements that can be utilized as freshly cultivated cells for rapid detection of nisin. The linear dose–response relationship perceived in the assay facilitates quantification of nisin in samples. The sensitivity of the nisin bioassay was 0.1 pg/ml in pure solution and 3 pg/ml in milk, exceeding the performance of all previously reported methods.