Fatih AKTAS, Elif GUDUK, Gulhan YASAR, Unzile GUVEN GULHAN

Soğuk Aktif ve Alkali Subtilisin Benzeri Proteaz Enziminin Bacillus sp. strain EL-GU1’ den İzolasyonu, Saflaştırılması ve Karakterizasyonu

Proteaz, amino asit rezidüleri arasındaki peptid bağlarını parçalayan ve deterjan, gıda, ilaç ve deri gibi çeşitli endüstriyel saha uygulamalarında kullanılan bir enzimdir. Proteaz enzim çeşitleri arasında endüstride en çok ilgi çeken, deterjan endüstrisinde ticari olarak en fazla potansiyel uygulama alanı bulan ve temizlik katkı maddesi olarak yaygın kullanılar alkalin proteazlardır.Bu çalışmada Bacillus sp. EL-GU1’den yeni bir alkali proteaz enzimi amonyum sülfat çöktürme metodu kullanılarak saflaştırıldı ve 16s rDNA dizileme yöntemi ile de proteaz olarak tanımlandı. En yüksek enzim aktivitesi, pH 6’da ve 25°C’de kazein substrat olarak kullanıldığında 3300 µmol/ min-1mg-1 olarak elde edildi. Sırasıyla enzim kinetik parametreleri olan KM, Vmax, kcat ve katalitik verimlilik değeri 1.4 mM, 1 mM/s, 2.10-7 s -1 , 0.14 10-7 s -1M -1 olarak hesaplandı. Bu sonuçlar bize gösteriyor ki Bacillus sp. ELGU1 suşu deterjan endüstrisinde kullanılmak için iyi bir aday olup ürettiği proteaz enzimi yeni ve özgün, düşük sıcaklıklarda da aktivite gösterebilen bir özelliktedir.

Isolation, Purification and Characterization of New Cold Active Subtilisin-like Protease from Bacillus sp. strain EL-GU1

One of the important hydrolytic enzymes are proteases that slice peptide bonds between amino acid residues.Proteases have various industrial applications including detergent, food, pharmaceutical, leather and diagnosticreagent industries. Among them, the most commercialized enzymes are alkaline proteases in the industry. Due totheir potential applications in the detergent industry as cleaning additives, they are of particular interest. In thisstudy, a novel protease from Bacillus sp. strain EL-GU1 was reported showing highest activity at pH 6 and20°C. The novel protease was purified by using ammonium sulfate precipitation and identified by 16S rDNAsequencing. Highest activity was observed as 3300 µmol/min-1mg-1 when casein used as a substrate. Kineticparameters of the enzyme were determined; KM, Vmax, kcat and catalytic efficiency values were calculated as 1.4mM, 1 mM/s, 2.10-7s-1, 0.14 10-7s-1M-1, respectively. These results indicated that the novel cold active proteasefrom Bacillus sp. strain EL- GU1 can be a good candidate for the detergent industry.

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