Detection of pediocin PA-1-producing pediococci by rapid molecular biology techniques |
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| Affiliation: | 1. Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, People’s Republic of China;2. Synergetic Innovation Center of Food Safety and Nutrition, Northeast Agricultural University, Harbin 150030, People’s Republic of China;3. College of Life Sciences, Jiamusi University, Jiamusi 154007, People’s Republic of China;1. Laboratory of Nano-Bioengineering, National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoye sh., 115409 Moscow, Russian Federation;2. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, 16/10 Miklukho-Maklaya, Russia;3. M.V. Lomonosov Moscow State University, 119991 Moscow, 1-12 Leninskie Gory, Russian Federation;4. Laboratoire de Recherche en Nanosciences, LRN-EA4682, Université de Reims Champagne-Ardenne, Reims 51100, France;1. Interdisciplinary Program for Bioengineering, Seoul National University, Seoul 151-744, South Korea;2. World Class University (WCU) Program of Chemical Convergence for Energy & Environment (C2E2), School of Chemical and Biological Engineering, College of Engineering, Seoul National University (SNU), Seoul 151-744, South Korea;3. School of Chemical and Biological Engineering, Seoul National University, Seoul 151-744, South Korea;4. Advanced Institutes of Convergence Technology, Suwon 443-270, South Korea |
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| Abstract: | Five bacteriocinogenic lactic acid bacteria strains (347, X13, Z102, A172 and P20) independently isolated from fermented sausages were identified asPediococcus acidilacticiby carbohydrate fermentation patterns and other biochemical characteristics. This fact, together with their activity againstListeria monocytogenes, suggested that they could be pediocin PA-1 producers. Rapid molecular biology techniques were used to detect the pediocin PA-1 operon in these strains. PCR, dot-blot and Southern hybridization, and DNA sequencing results confirmed that all of them had the genetic determinants for pediocin PA-1 biosynthesis encoded in a 9.4 kb plasmid. The bacteriocin produced byP. acidilactici347 has been purified by a procedure that included ammonium sulphate precipitation and cation exchange, hydrophobic-interaction and reverse-phase chromatography. The amino acid sequence of this bacteriocin was identical to pediocin PA-1, confirming the expression of the pediocin PA-1 genes. |
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