Isolation of Thermostable Alpha-amylase Producing Mesophilic Bacterium and Partial Characterization of the Enzyme

Abstract

Keywords

• Mesophilic Bacillus
• Isolation
• Alpha-amylase
• Thermal stability

Sıcaklığa Dirençli Alfa-amilaz Enzimi Üreten Mezofilik Bakteri İzolasyonu ve Enzimin Kısmi Karakterizasyonu

Öz

Bu çalışmada, Niğde ili Ulukışla ilçesi Çiftehan bölgesi sınırları içinde bulunan kaplıcadan alınan toprak örneklerinden α-amilaz aktivitesine sahip Bacillus izolasyonu yapılmıştır. İzolat bakteri, Bacillus sp. GA4 olarak isimlendirilmiştir. Bacillus sp. GA4 izolatına ait α-amilaz enziminin optimum pH ve sıcaklık değerleri sırasıyla 6.0-8.0 ve 50 ºC olarak bulunmuştur. Enzim 40 ºC’de 30 dk. ön inkübasyon sonrasında aktivitesini tamamen korumuştur. Enzim, 50 ºC ve 60 ºC’de 30 dk. ön inkübasyon sonrasında ise sırasıyla %21 ve %37’lik aktivite kaybına uğramıştır. MgCl2 ve FeSO4 enzim aktivitesini sırasıyla %13 ve %12 seviyelerinde artırırken, CuSO4, CaCl2 ve EDTA sırasıyla %81, %38 ve %33 oranlarında inhibe etmiştir. Bacillus sp. GA4 α-amilaz enziminin moleküler ağırlığı SDS-PAGE zimogram analizi ile yaklaşık olarak 55 kDa bulunmuştur. İzolatın CFX, CN, TE, RD, S, Amp ve P antibiyotiklerine karşı hassas, SH antibiyotiğine ise dirençli olduğu gözlenmiştir.

Anahtar Kelimeler

• Mezofilik Bacillus
• İzolasyon
• Alfa-amilaz
• Termal stabilite
• karakterizasyon

Kaynakça

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