İki ayrı lactococcus lactis alttür lactis suşundan bakteriyosin karakterizasyonu ve tanımlanması


Tezin Türü: Yüksek Lisans

Tezin Yürütüldüğü Kurum: Orta Doğu Teknik Üniversitesi, Türkiye

Tezin Onay Tarihi: 2004

Tezin Dili: İngilizce

Öğrenci: Oya Akçelik

Danışman: GÜLAY ÖZCENGİZ

Özet:

In this study, bacteriocins from two L. lactis subsp. lactis isolates of Turkey origin designated OC1 and OC2, respectively, were characterized and identified. The activity spectra of the bacteriocins were determined by using different indicator bacteria including Listeria, Bacillus and Staphylococcus spp. Bacteriocins were tested for their sensitivity to different enzymes, heat treatments and pH values. Loss of bacteriocin activities after ?-amylase treatment suggested that they form aggregates with carbohydrates. Molecular masses of partially purified bacteriocins were determined by SDS-polyacrylamide gel electrophoresis. PCR amplification was carried out with different primers for the detection of structural genes of lactococcal bacteriocins. As a result of these studies, the two bacteriocins were characterized as nisin and lacticin 481, respectively. Association of the bacteriocin production with plasmid DNA was examined by using acriflavine as a plasmid curing agent. Plasmid profiles of the wild type and its non-bacteriocin producing mutants were determined by using the alkali lysis method followed by agarose gel electrophoresis. The genetic nature of industrially important characteristics of Lactococcus lactis strains were investigated through gene transfer studies via conjugation. According to the results of plasmid curing and conjugal transfer trials, it was concluded that in Lactococcus lactis subsp. lactis OC1 strain a 39,7 kb plasmid is responsible for nisin production, lactose fermentation and proteolytic activity. In Lactococcus lactis subsp. lactis OC2 strain, on the other hand, a 16 kb plasmid appeared to be responsible for lacticin 481 production and lactose fermentation.