Purification and characterization of thermophilin T, a novel bacteriocin produced by Streptococcus thermophilus ACA-DC 0040 |
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| Authors: | A Aktypis G Kalantzopoulos JHJ Huis in't Veld & B ten Brink |
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| Institution: | Agricultural University of Athens, Department of Food Science and Technology, Athens, Greece,;University of Utrecht, Faculty of Veterinary Medicine, Department of the Science of Food of Animal Origin, Utrecht,;TNO Nutrition and Food Research, Department of Bioprocessing and Biomonitoring, Zeist, The Netherlands |
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| Abstract: | ACA-DC 0040 produced an antimicrobial agent, which was named thermophilin T, active against several lactic acid bacteria strains of different species and food spoilage bacteria, such as Clostridium sporogenes C22/10 and Cl. tyrobutyricum NCDO-1754. The crude antimicrobial compound is sensitive to proteolytic enzymes and α-amylase, heat-stable (100 °C for 30 min), resistant to pH exposure at pH 1–12 and demonstrates a bactericidal mode of action against the sensitive strain Lactococcus cremoris CNRZ-117. The production of bacteriocin was optimized approximately 10-fold in an aerobic fermenter held at constant pH 5·8 and 6·2. Ultrafiltration experiments with culture supernatant fluids containing the bacteriocin, and further estimation of molecular weight with gel filtration chromatography, revealed that bacteriocin in the native form has a molecular weight in excess of 300 kDa. SDS-gel electrophoresis of partially purified thermophilin T showed that bacteriocin activity was associated with a protein band of approximately 2·5 kDa molecular mass. |
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