Değişik inorganik taşıyıcılarda katalaz immobilizasyonu

Abstract

X Özet Çalışmamızda enzim immobilizasyonunda Si02-Aİ203/ perlit, aktif kum, kuartz inorganik taşıyıcı olarak kullanıldı. Bu inaktif taşıyıcılar 3-aminopropiltrietoksi- silan, Y~kloroProPİltrîetoksisixan' y-aminopropiltrime- toksisilan, ile aktive edilerek glutaraldehit ile çapraz bağlama yapıldı ve enzim kovalent olarak immobilize edildi. Taşıyıcıya kovalent olmayarak bağlanmış enzimler NaCİ içeren tampon ile yıkanarak uzaklaştırıldı ve elde edilen preparatlar vakum desikatöründe kurutularak aktivite tayinleri yapılmak üzere derin dondurucuda saklandı. Taşıyıcıya bağlanmış enzim miktarlarını saptamak amacı ile süzüntülerde Lowry yöntemine göre protein miktarı tayini yapıldı. Doğal ve bağlı enzimde sıcaklığa ve pH'a bağlı aktivite değişimleri araştırılarak optimum pH ve sıcaklık saptandı, immobilize enzimin işlem ve depolama kararlılıkları, inkübasyon süresinin aktivite üzerine etkisi incelenerek doğal enzimle kıyaslandı. Ayrıca doğal ve immobilize enzimde Fe3+, Ca2+, Mn2+ ve Mg2+ iyonlarının aktivite üzerine etkileri incelendi.

11 Abstract In this study, Si02, Sİ02-Aİ203, perlite, activated sand and quartz have been used as inorganic carriers in enzyme immobilization. Activation with 3-aminopropyltriethoxysilane, Y-cholopropyltr iethoxysilane, and with y- aminotrimethoxysilane was followed by cross-linking with glutar aldehyde, to this, enzyme was covalently immobilized. Enzyme which is not covalently bonded to matrices, was remowed by washing with buffer containing NaCl obtained products were dried in vacuum, and then kept in deep-freeze for further applications. In order to determine enzyme content of the filtrate, Lcwry method was used to find protein content, further enzymatic activity of natural and bonded enzyme was calculated respectively. Temperature and pH dependence of enzymatic activity were investigated, and optimum pH and temperature were probed. The effect of incubation period on the enzyme production and the period of activity time were determined. The effect of incubation period on enzyme activity, enzyme stability during production, and enzymatic shelf life were determined, and results compared with natural enzyme. Moreover, the content of milk, treated with the immobilized enzyme until the half life of the enzyme, was determined. And also Fe3+, Ca2+, Mn2+ and Mg2+ ion, effect on the natural and the immobilized enzyme activity were also investigated.