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A rapid turbidometric microplate bioassay for accurate quantification of lactic acid bacteria bacteriocins
Authors:Turcotte Carl  Lacroix Christophe  Kheadr Ehab  Grignon Louis  Fliss Ismaïl
Affiliation:

a Dairy Research Centre STELA, Pavillon Paul Comtois, Université Laval, Québec, PQ, Canada G1K7P4

b Laboratory of Food Biotechnology, Institute of Food Science and Nutrition, Swiss Federal Institute of Technology, ETH Zentrum, LFO F18 CH-8092, Zurich, Switzerland

c Department of Dairy Science and Technology, Faculty of Agriculture, University of Alexandria, Alexandria, Egypt

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 (r2=0.992±0.004) was obtained for the exponential regression in the nisin Z concentration range of 20–120 ng/ml with 1×107 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 r2=0.993±0.005, a precision (8.2%) and an accuracy lower than 15%.
Keywords:Nisin Z  Pediocin PA-1  Antimicrobial activity  Bioassay
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