CHARACTERIZATION OF BACTERIOCIN PRODUCED BY ANTILISTERIAL ENTEROCOCCUS MUNDTII YB6.30 ISOLATED FROM FERMENTED SUCUK

Öz

In this study, the characterization of bacteriocin produced by Enterococcus mundtii YB6.30 with antilisterial activity isolated from fermented sucuk was aimed. The YB6.30 inhibited various Gram-positive and Gram-negative bacteria, including Listeria species, as well as food pathogens such as Staphylococcus aureus, Bacillus cereus, Salmonella Enteritidis, S. Typhimurium and Escherichia coli. As a result of proteolytic enzyme treatment, the antimicrobial substance produced by YB6.30 was identified as bacteriocin. Identification of E. mundtii YB6.30 was done by 16S rRNA gene sequence analysis and polymerase chain reaction (PCR) using species-specific primers. As a result of different temperature and pH treatments, it was determined that bacteriocin is heat stable and shows activity in wide pH range (2.0-10.0). Bacteriocin showed a bactericidal activity against vancomycin resistant E. faecium ATCC 51559. As a result of the PCR experiment, the presence of mundtisin KS gene was detected in E. mundtii YB6.30. Molecular size of active protein band was determined as 7.56 kDa as a result of Tricine-SDS-PAGE analysis.

Anahtar Kelimeler

• Enterococcus mundtii
• mundtisin KS
• sucuk (Turkish dry-fermented sausage)
• enterocin
• polymerase chain reaction (PCR)
• Tricine-SDS-PAGE

FERMENTE SUCUKTAN İZOLE EDİLEN ANTİLİSTERİAL ENTEROCOCCUS MUNDTII YB6.30 TARAFINDAN ÜRETİLEN BAKTERİYOSİNİN KARAKTERİZASYONU

Öz

Bu çalışmada, fermente sucuktan izole edilen antilisterial aktiviteye sahip Enterococcus mundtii YB6.30 tarafından üretilen bakteriyosinin karakterizasyonu amaçlanmıştır. YB6.30 izolatı Listeria türlerinin yanı sıra Staphylococcus aureus, Bacillus cereus, Salmonella Enteritidis, S. Typhimurium ve Escherichia coli gibi gıda patojenlerinin de dahil olduğu çeşitli Gram-pozitif ve Gram-negatif bakterileri inhibe etmiştir. Proteolitik enzim uygulaması sonucu YB6.30 izolatı tarafından üretilen antimikrobiyal maddenin bakteriyosin olduğu belirlenmiştir. E. mundtii YB6.30 suşunun tanısı 16S rRNA gen dizi analizi ve türe özgü primerler kullanılarak yapılan polimeraz zincir reaksiyonu (PZR) ile yapılmıştır. Farklı sıcaklık ve pH uygulamaları sonucu bakteriyosinin ısı stabil olduğu ve geniş pH aralığında (2.0-10.0) aktivite gösterdiği belirlenmiştir. Bakteriyosin vankomisin dirençli E. faecium ATCC 51559 suşuna karşı bakterisidal etki göstermiştir. PZR denemesi sonucu E. mundtii YB6.30’da mundtisin KS geni varlığı tespit edilmiştir. Trisin-SDS-PAGE analizi sonucu aktif protein bandının moleküler büyüklüğü yaklaşık 7.56 kDa olarak tespit edilmiştir.

Anahtar Kelimeler

• Enterococcus mundtii
• mundtisin KS
• fermente sucuk
• enterosin
• polimeraz zincir reaksiyonu (PZR)
• Trisin-SDS-PAGE

Kaynakça

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