Seyma SENSOY KARAOGLU, Ali SEVİM, Elif SEVİM

Topraktan izole edilen Bacillus amyloliquefaciens B10 morfolojik, biyokimyasal ve 16S rRNA sekans analizi sonuçlarına göre belirlenmiştir. Bacillus amyloliquefaciens B10’un ürettiği bakteriyosin benzeri peptit (BLP) Bacillus cereus ve Listeria monoctogenesis gibi patojenik Gram pozitif ve Candida albicans gibi fırsatçı patojenleri içeren mayalara karşı geniş antimikrobiyal aktivite gösterir. BLP katalaz hariç proteolitik proteinaz-K, lipaz, tripsin, α-amilaz enzimlerine hassastır (5 mg/ml). BLP aktivitesi 90°C’de 30dk. da stabil iken aktivite 90°C 2 saatde ve 121°C’de 20 dk. da kaybolmaktadır. BLP pH 3-9 arasında aktivite göstermesine rağmen, optimum aktivite pH 7’dir. BLP ethanol, hekzan, etil eter ve tween-80 gibi bazı organik çözücü ve deterjanlara karşı hassastır. Amonyum sülfat çöktürmesi (%70) ile yapılan kısmi saflaştırma sonucu BLP’nin yaklaşik 5 kDa ağırlığında olduğu tespit edilmiştir.

Anahtar Kelimeler:

Bacillus amyloliquefaciens B10, bakteriyosin benzeri peptit, antimikrobiyal aktivite, kısmi saflaştırma

Production and Characterization of Bacteriocin-like Peptide Produced by Bacillus amyloliquefaciens B10

Bacillus amyloliquefaciens B10 that was isolated from soil was identified according to data of morphological, biochemical and 16S rRNA sequence analyses. The bacteriocin like peptide (BLP) produced by B. amyloliquefaciens B10 shows extendend antimicrobial activity against gram-positive bacteria including some pathogenic bacteria such as B. cereus and L. monocytogenesis and yeast including opportunistic pathogen such as Candida albicans. The BLP was sensitive to proteolytic enzyme proteinase K, lipase, tripsin, α- amylase except for catalase (5 mg/ml). While the activity of BLP of B13 was stable at 90°C for 30 min, the activity was completely lost at 90°C for 2 h and 121°C 20 min. Although activity of BLP of B10 was stable pH range 3-9, optimum pH was 7.0. It was sensitive against some organic solvent and detergent such as ethanol, hexane, ethyl ether and tween-80. Partial purification of BLP of B10 performed 70% ammonium sulphate precipitation was shown that it has approximately 5 kDa molecular weight.

Keywords:

Bacillus amyloliquefaciens B10, bacteriocin like peptide, antimicrobial activity, partial purification,

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