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A rapid turbidometric microplate bioassay for accurate quantification of lactic acid bacteria bacteriocins
Carl Turcotte1, Christophe Lacroix, Ehab Kheadr
1Dairy Research Centre STELA, Pavillon Paul Comtois, Université Laval, Québec, PQ, Canada G1K7P4.
Abstract:
A 1 day turbidometric microplate bioassay (TMB) was developed for the rapid, accurate and precise quantification of lactic acid bacteria (LAB) bacteriocins (nisin Z and pediocin PA-1). Parameters such as the concentration of the indicator strains and the incubation time were optimized for each bacteriocin. A high correlation coefficient (r(2)=0.992+/-0.004) was obtained for the exponential regression in the nisin Z concentration range of 20-120 ng/ml with 1 x 10(7) CFU indicator strain (Pediococcus acidilactici UL5) and an incubation time of 3 h. Using these parameters, the detection limit was estimated at 80 ng/ml (3.2 IU/ml), compared to 300 ng/ml for the agar diffusion assay (ADA). High precision (<7%) and accuracy (10%) were obtained for all nisin Z concentrations tested. Similar results were obtained with pediocin PA-1 with r(2)=0.993+/-0.005, a precision (8.2%) and an accuracy lower than 15%.

