Cu2+ takılı O-karboksimetil kitosan Schiff bazı kompleksleri gömülü kompozit membranların hazırlanması ve protein adsorpsiyon davranışlarının incelenmesi
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Dosyalar
Tarih
2015
Yazarlar
Dergi Başlığı
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Yayıncı
Aksaray Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışma kapsamında, lizozimin etkin bir şekilde ayrılması için afinite kromatografisi yöntemleri arasında, son yıllarda yoğun bir uygulama alanı bulan, İmmobilize Metal Afinite Kromatografisi (İMAK) kullanılmıştır. Bu amaçla, suda çözünebilir O-karboksimetil kitosan Schiff bazı-4a kompleksleri sentezlenmiştir ve Cu2+ iyonu takılmıştır. Monomer olarak poli(2-hidroksietil metakrilat), çapraz bağlayıcı olarak N,N?-metilen-bis-akrilamid, başlatıcı/aktivatör olarak N,N,N´,N´- tetrametilen diamin (TEMED)/amonyum persülfat (APS) kullanılmış ve radikal kriyokopolimerizasyon yöntemiyle 5 mL'lik plastik tüplerde sentezlenmiştir. Hazırlanan Cu2+ takılı O-karboksimetil kitosan Schiff bazı kompleksleri gömülü kompozit kriyojel şişme testleri, taramalı elektron mikroskobu (SEM), FT-IR ile karakterize edilmiştir. Sulu çözeltiden lizozim adsorpsiyonuna pH'nın, başlangıç protein derişiminin, akış hızının, iyonik şiddetin ve sıcaklığın etkisi incelenmiştir. Maksimum adsorpsiyon pH 8 fosfat tamponunda, 3 mg/mL başlangıç lizozim derişiminde, 0,5 mL/dak akış hızında ve 25 oC ortam sıcaklığında 103,3 mg/g partikül olarak gerçekleşmiştir. Ayrıca, süpermakro-gözenekli kriyojellere proteinin defelarca adsorplanıp desorplanabildiği belirlenmiştir.
In this study, Immobilized Metal Affinity Chromatograph (IMAC) finding a wide application area among affinity chromatography methods was used for removal of lysozyme efficiently from aqueous solutions. For this purpose, water soluble Ocarboxymethyl chitosan complexes was synthesised, Cu2+ ions was attached onto them. The composite cryogels were prepared by free radical polymerization using N,N,N´,N´-tetramethylene diamine (TEMED) and ammonium persulfate (APS) as initiator/activator pair, N,N´-methylene-bis-acrylamide (MBAAm) as cross-linker in a plastic syringe of 5 mL. Prepared composit cryogel embedded with Cu2+ -attached Ocarboxymethyl chitosan Schiff base complexes was characterized by FT-IR, SEM, swelling tests measurements. The effect of pH, ionic strength, initial lysozyme concentration, temparature and flow rate on adsorption were investigated. The maximum amount of lysozyme adsorption from aqueous solution was at 103,3 mg/g particles in phosphate buffer at pH 8 with initial lysozyme concentration of 3 mg/mL. On the other hand, the most efficent adsorption was carried out at 0,5 mL/min, at 25oC temparature. It was also observed that enzyme could be repeatedly adsorbed and desorbed on the Cu2+ attached O-carboxymethyl chitosan Schiff base complexes croygels.
In this study, Immobilized Metal Affinity Chromatograph (IMAC) finding a wide application area among affinity chromatography methods was used for removal of lysozyme efficiently from aqueous solutions. For this purpose, water soluble Ocarboxymethyl chitosan complexes was synthesised, Cu2+ ions was attached onto them. The composite cryogels were prepared by free radical polymerization using N,N,N´,N´-tetramethylene diamine (TEMED) and ammonium persulfate (APS) as initiator/activator pair, N,N´-methylene-bis-acrylamide (MBAAm) as cross-linker in a plastic syringe of 5 mL. Prepared composit cryogel embedded with Cu2+ -attached Ocarboxymethyl chitosan Schiff base complexes was characterized by FT-IR, SEM, swelling tests measurements. The effect of pH, ionic strength, initial lysozyme concentration, temparature and flow rate on adsorption were investigated. The maximum amount of lysozyme adsorption from aqueous solution was at 103,3 mg/g particles in phosphate buffer at pH 8 with initial lysozyme concentration of 3 mg/mL. On the other hand, the most efficent adsorption was carried out at 0,5 mL/min, at 25oC temparature. It was also observed that enzyme could be repeatedly adsorbed and desorbed on the Cu2+ attached O-carboxymethyl chitosan Schiff base complexes croygels.
Açıklama
Anahtar Kelimeler
Afinite, Lizozim, Kriyojel, İMAK, Karboksimetil Kitosan, Adsorpsiyon, Lysozyme, Cryogel, Affinity, IMAC, Carboxymethyl Chitosan
