Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T
Yıl: 2015 Cilt: 40 Sayı: 2 Sayfa Aralığı: 116 - 124 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022
Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T
Öz: Amaç: Bu çalışmanın amacı termofilik bir bakteri olan Thermus sp. NCCB 100425T'den bir esteraz enziminin saflaştırılıp karakterize edilmesi ve endüstriyel uygulamalar için uygunluğunun incelenmesidir.Metod: Thermus sp. NCCB 100425T esterazı amonyum sülfat çöktürmesi ve hidrofobik etkileşim kromatografisi kullanılarak saflaştırıldı ve biyokimyasal özellikleri incelendi.Bulgular: Doğal- ve SDS-PAGE'de tek bant şeklinde görülen saf enzimin, p-nitrofenil asetat (pNPA) varlığında optimum pH ve sıcaklık değerleri sırasıyla 7,5 ve 60°C olarak belirlendi. Yine pNPA bir substrat olarak kullanıldığında Km ve Vmax değerlerinin 18,32 mM ve 96,15 U/mg protein olduğu tespit edildi. pH kararlılığı pH 4,0-9,0 aralığında 4°C ve 60°C'de incelendi. 7 günlük bir inkübasyondan sonra saf enzimin aktivitesinin 4°C'de bütün pH'larda yaklaşık %85-90 oranında korunduğu, bu oranın 60°C'de pH 8,0'de ise %59±5,2 olduğu görüldü. 30-60°C'ler arasında 7 gün inkübe edilen enzim, orijinal aktivitesini %80 oranında korudu. %10 nihai konsantrasyondaki etanol ve DMSO varlığında enzim aktivitesi sırasıyla %96±2,7 ve %78±2,5 oranlarında korundu. Ayrıca, bazı metal iyonlarının enzim aktivitesini farklı oranlarda etkilediği tespit edildi. Sonuç: Bütün bu sonuçlar değerlendirildiğinde Thermus sp. NCCB 100425T esterazının özellikle yüksek sıcaklık- ve pH-kararlılığı nedeniyle endüstriyel ve/veya klinik uygulamalar için avantajlara sahip olabileceği açıkça görülmektedir.
Anahtar Kelime: Konular:
[Thermus sp. NCCB 100425T'den yeni bir ısıya dayanıklı esterazın saflaştırılması ve karakterizasyonu]
Öz: Objective: The purpose of the present study was to purify and characterize an esterase from a thermophilic bacterium Thermus sp. NCCB 100425T and to check its suitability for industrial applications.Methods: Thermus sp. NCCB 100425T esterase was purified by using ammonium sulphate precipitation and hydrophobic interaction chromatography and then characterized biochemically.Results: The purity of the enzyme was observed as a single band on native- and SDS- PAGE. In the presence of p-nitrophenyl acetate (pNPA) as a substrate, the optimum pH and temperature of the enzyme were found to be 7.5 and 60°C, respectively. Km and Vmax values are calculated as 18.32 mM and 96.15 U/mg protein, respectively, with pNPA. pH stability was investigated in the range of pH 4.0-9.0 at 4°C and 60°C. After 7 days incubation, activity of pure enzyme was retained 85-90% for all pH at 4°C and 59±5.2% for pH 8.0 at 60°C. It was determined that approximately 80% of enzyme activity was retained between 30-60°C after 7 days incubation. In the presence of 10% ethanol and DMSO, the enzyme activity was retained 96±2.7% and 78±2.5%, respectively. Additionally, it was detected that some metal ions affect the enzyme activity at different ratios.Conclusion: It is clear that Thermus sp. NCCB 100425T esterase might have advantages for industrial and/or clinical applications in terms of especially its high thermal- and pH-stability.
Anahtar Kelime: Konular:
Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık
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| APA | TEKİNCANLİ M, YILDIRIM AKATIN M, Çolak A (2015). Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. , 116 - 124. |
| Chicago | TEKİNCANLİ Mehmet Ali,YILDIRIM AKATIN Melike,Çolak Ahmet Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. (2015): 116 - 124. |
| MLA | TEKİNCANLİ Mehmet Ali,YILDIRIM AKATIN Melike,Çolak Ahmet Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. , 2015, ss.116 - 124. |
| AMA | TEKİNCANLİ M,YILDIRIM AKATIN M,Çolak A Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. . 2015; 116 - 124. |
| Vancouver | TEKİNCANLİ M,YILDIRIM AKATIN M,Çolak A Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. . 2015; 116 - 124. |
| IEEE | TEKİNCANLİ M,YILDIRIM AKATIN M,Çolak A "Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T." , ss.116 - 124, 2015. |
| ISNAD | TEKİNCANLİ, Mehmet Ali vd. "Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T". (2015), 116-124. |
| APA | TEKİNCANLİ M, YILDIRIM AKATIN M, Çolak A (2015). Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. Türk Biyokimya Dergisi, 40(2), 116 - 124. |
| Chicago | TEKİNCANLİ Mehmet Ali,YILDIRIM AKATIN Melike,Çolak Ahmet Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. Türk Biyokimya Dergisi 40, no.2 (2015): 116 - 124. |
| MLA | TEKİNCANLİ Mehmet Ali,YILDIRIM AKATIN Melike,Çolak Ahmet Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. Türk Biyokimya Dergisi, vol.40, no.2, 2015, ss.116 - 124. |
| AMA | TEKİNCANLİ M,YILDIRIM AKATIN M,Çolak A Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. Türk Biyokimya Dergisi. 2015; 40(2): 116 - 124. |
| Vancouver | TEKİNCANLİ M,YILDIRIM AKATIN M,Çolak A Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T. Türk Biyokimya Dergisi. 2015; 40(2): 116 - 124. |
| IEEE | TEKİNCANLİ M,YILDIRIM AKATIN M,Çolak A "Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T." Türk Biyokimya Dergisi, 40, ss.116 - 124, 2015. |
| ISNAD | TEKİNCANLİ, Mehmet Ali vd. "Purification and characterization of a novel thermostable esterase from Thermus sp. NCCB 100425T". Türk Biyokimya Dergisi 40/2 (2015), 116-124. |
