Alfa-amilaz ve gama-amilaz enzimlerinin çapraz bağlı enzim agregatları (CLEA) şeklinde immobilizasyonu
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Tarih
2013
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Çanakkale Onsekiz Mart Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, üç farklı amilaz enziminin çapraz bağlı enzim agregatları biçiminde immobilizasyonu gerçekleştirildi. Bu amaçla Aspergillus oryzea ve Bacillus licheniformis alfa - amilaz enzimleri ile Aspergillus niger gama - amilaz enzimi kullanıldı. Enzimler 1,2 - dimetoksietan, amonyum sülfat, aseton, dimetilsülfoksit ve etanol kullanılarak agregatlaştırıldı. Elde edilen agregatların glutaraldehit ile çapraz bağlanması sonucunda çözünmez formda, katalitik olarak aktif enzim agregatları elde edildi. Belirli optimizasyon koşulları altında Aspergillus oryzea alfa - amilazı, Bacillus licheniformis alfa - amilazı ve Aspergillus niger gama - amilazı için sırasıyla % 21,8; % 94,1 ve % 41,2 aktivite geri kazanımları gözlendi. Aspergillus oryzea alfa - amilazı ve Aspergillus niger gama - amilazı için aktivite geri kazanımı değerlerinin hedeflenenden düşük bulunması sebebiyle, bu enzimlerin çapraz bağlanmasında glutaraldehit yerine dekstran polialdehit de denendi. Bu amaçla glutaraldehit için optimize edilen koşullarda farklı dekstran polialdehit miktarları ile çapraz bağlama reaksiyonları gerçekleştirildi. Bunun sonucunda Aspergillus oryzea alfa - amilazı için % 60,0 aktivite geri kazanımı gözlendi; Aspergillus niger gama - amilaz enzimi için ise herhangi bir aktivite geri kazanımı gözlenemedi. Elde edilen Bacillus licheniformis alfa - amilazı çapraz bağlı enzim agregatlarına ait bazı kinetik parametreler hesaplandı. Anahtar Sözcükler: Enzim immobilizasyonu, çapraz bağlı enzim agregatları, amilaz, glukoamilaz, glutaraldehit, dekstran polialdehit
In this study, three different amylase enzymes were immobilized as their cross - linked enzyme aggregates. For this purpose Aspergillus oryzea and Bacillus licheniformis alpha - amylases and Aspergillus niger gamma - amylase were used. 1, 2 - dimethoxyethane, ammonium sulfate, acetone, dimethylsulphoxide and ethanol were used as precipitants. As a result of the cross-linking of resulting insoluble aggregates with glutaraldehyde, catalytically active enzyme aggregates were obtained. Under certain optimization conditions, activity recovery of 21,8 %; 94,1 % and 41,2 % was obtained by the cross - linked enzyme aggregates of Aspergillus oryzea alpha - amylase, Bacillus licheniformis alpha - amylase and Aspergillus niger gamma - amylase respectively. Due to the low activity recovery values for Aspergillus oryzea alpha - amylase and Aspergillus niger gamma - amylase, dextran polyaldehyde was also investigated as a cross - linker instead of glutaraldehyde. For this purpose, cross - linking reactions with different amounts of dextran polyaldehyde were carried out with the same conditions which are optimized for glutaraldehyde. As a result, activity recovery of 60,0 % was obtained for Aspergillus oryzea alpha - amylase. On the other hand no activity recovery was observed for Aspergillus niger gamma - amylase. Some kinetic parameters of resulting cross - linked enzyme aggregates of Bacillus licheniformis alpha - amylase were calculated. Keywords: Enzyme immobilization, cross - linked enzyme aggregates, amylase, glucoamylase, glutaraldehyde, dextran polyaldehyde
In this study, three different amylase enzymes were immobilized as their cross - linked enzyme aggregates. For this purpose Aspergillus oryzea and Bacillus licheniformis alpha - amylases and Aspergillus niger gamma - amylase were used. 1, 2 - dimethoxyethane, ammonium sulfate, acetone, dimethylsulphoxide and ethanol were used as precipitants. As a result of the cross-linking of resulting insoluble aggregates with glutaraldehyde, catalytically active enzyme aggregates were obtained. Under certain optimization conditions, activity recovery of 21,8 %; 94,1 % and 41,2 % was obtained by the cross - linked enzyme aggregates of Aspergillus oryzea alpha - amylase, Bacillus licheniformis alpha - amylase and Aspergillus niger gamma - amylase respectively. Due to the low activity recovery values for Aspergillus oryzea alpha - amylase and Aspergillus niger gamma - amylase, dextran polyaldehyde was also investigated as a cross - linker instead of glutaraldehyde. For this purpose, cross - linking reactions with different amounts of dextran polyaldehyde were carried out with the same conditions which are optimized for glutaraldehyde. As a result, activity recovery of 60,0 % was obtained for Aspergillus oryzea alpha - amylase. On the other hand no activity recovery was observed for Aspergillus niger gamma - amylase. Some kinetic parameters of resulting cross - linked enzyme aggregates of Bacillus licheniformis alpha - amylase were calculated. Keywords: Enzyme immobilization, cross - linked enzyme aggregates, amylase, glucoamylase, glutaraldehyde, dextran polyaldehyde
Açıklama
Fen Bilimleri Enstitüsü, Kimya Ana Bilim Dalı
Anahtar Kelimeler
Biyokimya, Biochemistry
