Tr. J. of Biology 22 (1998) 217-228 © TÜBİTAK
Purification and Characterization of Pediocin F, A Bacteriocin Produced By Pediococcus acidilactici F
Gazi University, Faculty of Arts and Science, Department of Biology Teknikokullar, 0650 Ankara -TURKEY
Middle East Technical University, Faculty of Arts and Science, Department of Biology, Ankara-TURKEY
Gazi University, Faculty of Arts and Science, Department of Biology, Teknikokullar, 06500 Ankara-TURKEY
Ankara University, Faculty of Science, Department of Biology, 06100 Ankara-TURKEY
Abstract: In this work, Pediococcus acidilactici F isolated from fermented sausage was reported to produce an extracellular antimicrobial peptide, termedPediocin F, that was protein in nature and effective against many bacteria associated with food spoilage and food related health hazards. Pediocin F is reported here to be sensitive to proteolytic enzymes, resistant to heat and organic solvents, and active over a wide range of pH. We have also reported that the extracellular antimicrobial protein produced by P. acidilactici F was really a smallpeptide. Coomassie-blue stained SDS-PAGE technique allowed us to estimate the molecular weight of this small peptide to be about 4460 dalton. Key Words: Pediococcus acidilactici, Pediocin, characterization, SDS-PAGE
Pediococcus acidilactici F Tarafından Üretilen Pediocin F’ in Saflaştırılıp Karakterize Edilmesi
Özet: Bu çalışmada fermente sosisden izole edilen Pediococcus acidilactici F’ in proteinyapısında olan, gıdalarla ilgili sağlığı tehtit eden ve gıdaların bozulmasına sebep olan bir çok bakteriye karşı Pediocin F olarak adlandırılan hücre dışı bir antimikrobiyal peptit ürettiği rapor edilmiştir. Pediocin F, proteolitik enzimlere karşı hassas, ısıya ve organik çözücülere karşı dayanıklı ve geniş bir pH skalasında aktiftirler. Coomassie-blue ile boyanmış SDS-PAGE tekniği bakterininantimikrobiyal etkisi ile ilgili protein bandının tanımlanması amacı ile kullanılmış ve bu çalışma sonucunda acidilactici F tarafından üretilen ve Pediocin F olarak adlandırılan bu proteinin 4460 dalton moleküler ağırlığına sahip oldukça küçük bir peptit olduğu bulunmuştur. Anahtar Sözcükler: Pediococcus acidilactici, Pediocin, karakterizasyon, SDS-PAGE
Purification and Characterization ofPediocin F, A Bacteriocin Produced by Pediococcus acidilactici F
Introduction Several species and strains of Pediococci have been used as starter cultures in the fermentation of vegetables (26), meats (9,28), sausage products (29) and cheddar cheese (11). Pediococci are lactic acid bacteria which help to preserve fermented foods and contribute in flavor development (11, 22). Bacteriocins fromPediococcus species, designated as Pediocins, were shown to be protein in nature and inhibitory to several other bacteria, many of which are associated with food spoilage and health hazards of food origin (4, 7, 10, 12, 13, 15, 19, 24, 27, 32). Several unique properties such as activity over a wide pH range and after high- and lowtemperature treatment make them suitable as biological preservativesto extend the shelf-life of refrigerated semipreserved foods and canned foods (1, 21, 23). Some of these studies reported antimicrobial activity against different bacterial species, but little is known about the chemical and physical nature of these antimicrobial substrances. Gonzalez and Kunka (12) reported that P. acidilactici PAC 1.0 produced a bacteriocin, Pediocin PA1, which has a moleculerweight of 16500 daltons determined by gel-filtration and proteinaceous in nature as indicated by sensitivity to various proteolytic enzymes. Bhunia et al. (4) reported the proteinaceous nature of Pediocin AcH produced by P. acidilactici strain H and have also reported that this antimicrobial substance was effective against many bacteria associated with food spoilage and food related health...
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