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TARHANADAN İZOLE EDİLEN LAKTOBASİLLER TARAFINDAN ÜRETİLEN BAKTERİYOSİNLERİN KARAKTERİZASYONU

Tarhana Anadolu’da kış için hazırlanan ve sık tüketilen fermente bir gıdadır. Fermente tarhana hamuru laktik asit bakterileri (LAB) ve maya türlerinden oluşan mikrofloraya sahiptir. Bu florada bazı LAB’de bakteriyosin üretimiyle antimikrobiyal aktivite gösterirler. Çalışmamızın amacı tarhanadan izole edilmiş L. namurensis PFC70, L. plantarum PFC74 ve L. paralimentarius PFC97 suşlarının bakteriyosinlerinin belirlenmesi ve karakterizasyonudur. PFC70, PFC74 ve PFC97 suşlarının Micrococcus luteus DSM1790 suşuna karşı 400, 1600, 1600 AU/mL antimikrobiyal aktiviteye sahip olduğu belirlenmiştir. Suşların kültür üst sıvılarındaki metabolitlerin yüksek sıcaklığa ve proteaz enzimlerine karşı hassas, düşük pH koşullarında stabil, bakteriyosin tabiatında olduğu anlaşılmıştır. Üretici hücrelerin genomunda yapılan PZR taramasında, PFC74 bakteriyosininin plantarisin benzeri olduğu belirlenmiştir. Bakteriyosinler, amonyum sülfat çöktürmesi, katı faz ekstraksiyonu ve ters faz sıvı kromotografisi ile saflaştırılmış ve trisin-SDS PAGE ile moleküler büyüklükleri 5 kDa altında olduğu tespit edilmiştir. PFC70 ve PFC74 bakteriyosini bakteriyosidal, PFC97 ise bakteriyostatik etkili bulunmuştur. Bu sonuçlar PFC70 suşunun yeni bir bakteriyosin üreticisi olduğunu göstermiştir.

Anahtar Kelimeler:

Tarhana, Laktik Asit Bakterisi, Bakteriyosin, Antimikrobiyal

CHARACTERIZATION OF BACTERIOCINS PRODUCED BY LACTOBACILLI ISOLATED FROM TARHANA

Tarhana is a fermented food that has been consumed often, produced for winter in Anatolia. Fermented tarhana dough microflora includes lactic acid bacteria (LAB), yeast species and some LAB exhibit antimicrobial activity due to bacteriocin production. The purpose of study was determination and characterization of bacteriocins of L. namurensis PFC70, L. plantarum PFC74, L. paralimentarius PFC97 strains which were isolated from tarhana. Strains had 400, 1600, 1600 AU/mL antimicrobial activity against Micrococcus luteus DSM1790 respectively. The relevant methabolites of strains at the culture supernatant was to be bacteriocin nature with low pH stability, high temparature and protease enzymes sensitivity. Producer cell genome showed that PFC74 bacteriocin was similiar with plantaricin. Bacteriocins were purified by ASP, SPE, RPLC and thereby molecular weights were determined under 5 kDa with Tricine-SDS-PAGE. PFC70 and PFC74 bacteriocin were found to be bacteriocidal, while PFC97 was bacteriostatic. Results indicated that PFC70 was novel bacteriocin producer.

Keywords:

Tarhana, Lactic acid bacteria, bacteriocin, antimicrobial,

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