Publication: a-Amilaz Enziminin Fotoimmobilizasyonu
Abstract
α–AMİLAZ ENZİMİNİN FOTOİMMOBİLİZASYONU Bu çalışmada, UV ile sertleştirilebilen N-(4-sodyumsülfofenil)-maleimid monomeri sentezlenerek enzim bağlama potansiyeli incelendi. Sentezlenen monomer, kovalent bağlanma için Brom ile aktifleştirilerek, enzimin yüzeyindeki gruplardan ılıman şartlarda (0 oC, 30 dakika) reaksiyon vermesine olanak sağlandı. Bu yöntemde, sülfonil bromür uçlu monomer proteinin amino gruplarıyla reaksiyona girerek, sülfonamid bağları oluşturdu. Polimerik destek UV-ışınları yardımıyla sertleştirme tekniğiyle hazırlandı. Immobilizasyon verimi 68,18 mg/ g monomer olarak bulundu. Maksimum aktivite pH 6,5’ta gözlemlendi. Immobilizasyon, pH’a bağımlı olan enzim aktivitesini değişmemiştir. Serbest enzim için optimum sıcaklık yaklaşık 30 ˚C bulunmuşken, immobilize enzim için yaklaşık 50 ˚C’ye kaymıştır. Serbest enzim 15 gün içinde aktivitesini kaybetmiştir. İmmobilize enzim ise 30 gün içinde aktivitesinin sadece % 30’nu kaybetmiştir.
PHOTOIMMOBILIZATON OF α –AMYLASE ENZYME In this study, UV-curable N-(4-Sodiumsulfophenyl)-maleimide monomer was synthesized and its potential for enzyme binding was investigated. The Bromine, which is used to activate the synthesized monomer for covalent attachments, has the advantage of giving reaction with the surface groups of enzyme under very mild conditions (0 oC, 30 minute). In this procedure, sulfonyl bromide pendant monomer reacted with amino groups of the protein to form sulfonamide bonds. Polymeric support was prepaped by UV-curing technique. The Immobilization yield was found to be as 68.18 mg/ g monomer. The maximum activity was observed at pH 6.5. Immobilization did not change the pH-dependency of the enzyme activity. It was found that the optimum temperature for the free enzyme was approximately 30 oC, while it shifted to nearly 50 oC for the immobilized enzyme. Free enzyme lost its activity completely within 15 days. Immobilized enzyme lost only 30 % of its activity in 30 days.
PHOTOIMMOBILIZATON OF α –AMYLASE ENZYME In this study, UV-curable N-(4-Sodiumsulfophenyl)-maleimide monomer was synthesized and its potential for enzyme binding was investigated. The Bromine, which is used to activate the synthesized monomer for covalent attachments, has the advantage of giving reaction with the surface groups of enzyme under very mild conditions (0 oC, 30 minute). In this procedure, sulfonyl bromide pendant monomer reacted with amino groups of the protein to form sulfonamide bonds. Polymeric support was prepaped by UV-curing technique. The Immobilization yield was found to be as 68.18 mg/ g monomer. The maximum activity was observed at pH 6.5. Immobilization did not change the pH-dependency of the enzyme activity. It was found that the optimum temperature for the free enzyme was approximately 30 oC, while it shifted to nearly 50 oC for the immobilized enzyme. Free enzyme lost its activity completely within 15 days. Immobilized enzyme lost only 30 % of its activity in 30 days.