Sığır karaciğeri kaynaklı katalaz enziminin kalsiyum alginat jele immobilizasyonu
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Dosyalar
Tarih
2012
Yazarlar
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Yayıncı
Hatay Mustafa Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada sığır karaciğer katalazının (CAT) kalsiyum alginat jele(Ca-Alg) tutuklama yöntemi ile immobilizasyonu araştırılmıştır. İmmobilizasyon koşullarının optimizasyonu yapıldığında en yüksek aktivite başlangıç enzim derişimi 1 mg/mL, Na-Alg derişimi % 2 ve CaCl2 derişimi 1,5 M kullanıldığında elde edilmiştir. Kalsiyum alginat; immobilizasyon sırasında glukoz, kitosan ve diaminohekzan (DAH) ile modifiye edilmiştir. En yüksek aktiviteyi gösteren DAH+ Ca-Alg/CAT örneğinin özellikleri, modifiye edilmemiş Ca-Alg jele immobilize CAT (Ca-Alg/CAT) ile kıyaslanmıştır.Serbest CAT, Ca-Alg/CAT ve DAH+Ca-Alg/CAT örneklerinin maksimum aktivite gösterdiği pH ve sıcaklık değerleri sırasıyla 7,5, 30 ? C- 8,0, 45 ? C ve 8,0, 40 ? C olarak belirlenmiştir. Belirlenen optimum koşullarda Ca-Alg/CAT ve DAH+ Ca-Alg/CAT örneklerinin Vmax değeri serbest enzimin aktivitesinin sırasıyla % 6,27 ve % 7,36'sını göstermiştir. 24 saat süreyle termal kararlılıklar incelenmiş, serbest CAT'ın 30 ? C'de % 45 Ca-Alg/CAT'ın 45 ? C'de % 60 ve DAH+ Ca-Alg/CAT'ın 40 ? C % 74 aktivitesinin kaldığı belirlenmiştir. Ca-Alg/CAT ve DAH+ Ca-Alg/CAT örneklerinin 4 ? C ve oda sıcaklığında depolama kararlılıkları incelenmiş, 40 gün sonunda Ca-Alg/CAT ve DAH+Ca-Alg/CAT örneklerinin kalan aktiviteleri sırasıyla 4 ? C'de, % 45,2 ve % 59,5; oda sıcaklığında ise % 13 ve % 25,1 olarak belirlenmiştir. Ca-Alg/CAT ve DAH+Ca-Alg/CAT sürekli reaktörde kullanımı da araştırılmış ve DAH+Ca-Alg/CAT örneğinin kullanım için daha uygun olduğu belirlenmiştir.
In this study, immobilization of bovine liver catalase (CAT) into calcium alginat beads by entrapment was investigated. In the optimization studies of immobilization, the highest activity was obtained when Na-Alg, CaCl2 and initial enzyme concentration were used as 2%, 1.5 M, 1 mg/mL, respectively. Calcium alginate (Ca-Alg) was modified with glucose, chitosan and diaminohexane(DAH). DAH+ Ca-Alg/CAT showed the best activity and its properties was compared with immobolized CAT with unmodified Ca-Alg gel (Ca-Alg/CAT).Optimum pH and temperature values of free CAT, Ca-Alg/CAT and DAH+Ca-Alg/CAT were determined as 7.5, 30 ? C- 8.0, 45 ? C ve 8.0, 40 ? C, respectively. The Vmax values of Ca-Alg/CAT and DAH+Ca-Alg/CAT at optimal conditions were about 6.27% and 7.36% of free CAT, respectively. Thermal stabilities of free CAT, Ca-Alg/CAT and DAH+Ca-Alg/CAT were investigated and they retained about 45% when preincubated at 30 ? C, 60% at 45 ? C and 74% at 40 ? C of their initial activities for 24 h, respectively, Ca-Alg/CAT and DAH+ Ca-Alg/CAT retained 45.2% and 59.5% of their initial activities at 4 ? C; 13% and 25.1% of their initial activities at room temperature, respectively at the end of 40 days. Operational stabilities of Ca-Alg/CAT and DAH+ Ca-Alg/CAT were investigated in plug- ? ow type reactor and it was observed that DAH+ Ca-Alg/CAT sample was more proper to use.
In this study, immobilization of bovine liver catalase (CAT) into calcium alginat beads by entrapment was investigated. In the optimization studies of immobilization, the highest activity was obtained when Na-Alg, CaCl2 and initial enzyme concentration were used as 2%, 1.5 M, 1 mg/mL, respectively. Calcium alginate (Ca-Alg) was modified with glucose, chitosan and diaminohexane(DAH). DAH+ Ca-Alg/CAT showed the best activity and its properties was compared with immobolized CAT with unmodified Ca-Alg gel (Ca-Alg/CAT).Optimum pH and temperature values of free CAT, Ca-Alg/CAT and DAH+Ca-Alg/CAT were determined as 7.5, 30 ? C- 8.0, 45 ? C ve 8.0, 40 ? C, respectively. The Vmax values of Ca-Alg/CAT and DAH+Ca-Alg/CAT at optimal conditions were about 6.27% and 7.36% of free CAT, respectively. Thermal stabilities of free CAT, Ca-Alg/CAT and DAH+Ca-Alg/CAT were investigated and they retained about 45% when preincubated at 30 ? C, 60% at 45 ? C and 74% at 40 ? C of their initial activities for 24 h, respectively, Ca-Alg/CAT and DAH+ Ca-Alg/CAT retained 45.2% and 59.5% of their initial activities at 4 ? C; 13% and 25.1% of their initial activities at room temperature, respectively at the end of 40 days. Operational stabilities of Ca-Alg/CAT and DAH+ Ca-Alg/CAT were investigated in plug- ? ow type reactor and it was observed that DAH+ Ca-Alg/CAT sample was more proper to use.
Açıklama
Anahtar Kelimeler
Kimya, Chemistry
