Lizozim adsorpsiyonuna yönelik yeni bir kriyojelin hazırlanması ve enzim için bazı optimizasyon parametrelerinin belirlenmesi
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Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Dicle Üniversitesi, Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, 2-hidroksietil metakrilik asit (HEMA) ve metakriloil antipirin-Ce3+ (MAAP- Ce3+) kompleksinden yola çıkarak serbest radikal polimerizasyon yöntemi ile 2-hidroksietil metakrilik asit-metakriloil antipirin-Ce3+ [P(HEMA-MAAP-Ce3+)] kriyojeli hazırlandı. MAAP-Ce3+ kompleksinin UV-yakın infrared (UV-NIR) ve enerji dağıtıcı X-ray (EDX) teknikleri, p(HEMA-MAAP-Ce3+) kriyojelinin ise taramalı elektron mikroskobu (SEM), FT-IR ve şişme testleri ile karakterizasyon işlemi yapıldı. UV-NIR spektrumunda MAAP-Ce 3+ kompleksi için 916, 1037, 1081, 1109 ve 1141 nm'de beş farklı pik görüldü. EDX spektrumunda ise MAAP-Ce3+ kompleksinde %15,12 N ve %2,89 Ce3+ varlığı yapıda azot ve Ce3+ olduğunu gösterdi. FT-IR spektrumu çalışmalarında kriyojelin 3412 cm-1 'de –OH ve 3104 cm-1 'de –NH titreşimlerine, 1768-1712 cm-1 'de ise ester, amid ve siklik karbonil gruplarının C=O bağlarına sahip olduğu görüldü. Daha sonra kriyojel sulu çözeltiden lizozim adsorpsiyonu için kullanıldı. P(HEMA-MAAP-Ce3+) kriyojelin denge şişme oranı 7,39 g H2O/g kriyojel, kriyojelde oluşan makrogözenek miktarı ise % 80,8 olarak hesaplandı. Kriyojel gözenek boyutu ise 30-50 µm olarak bulundu. Lizozim adsorpsiyonu üzerine akış hızı, pH, sıcaklık, başlangıç enzim derişimi ve iyonik şiddetin etkisi incelendi. Maksimum adsorpsiyon kapasitesine (57,84 mg/g kriyojel) 0,5 mL/dak. akış hızında, pH: 6,0 ve 25 oC'de ulaşıldı. İyonik şiddetin etkisi sonucu, tuz derişiminin 0,05 M' den 1,0 M'ye artmasıyla lizozim adsorpsiyon kapasitesinin 41,31 mg/g'den 2,01 mg/g' ye kadar azaldığı tespit edildi. Langmuir ve Freundlich izoterm analizleri sonucu hesaplanan korelasyon katsayısı (R2 : 0,9939) Langmuir adsorpsiyon modelinin bu sisteme uygun olabileceğini gösterdi. Adsorplanan enzim, 20 mM, pH: 6,0 fosfat tamponu içeren 1 M NaCl çözeltisi ile 120 dak.'da %95,7 oranında desorbe edildi. Tekrar kullanılabilirlik çalışmalarında, kriyojelin 7 kez ard arda kullanılmasına rağmen adsorpsiyon kapasitesinde fazla bir düşüş olmadığı görüldü.
n this study, 2-hydroxyethyl methacrylic acid-methacryloyl antipyrin-Ce3+[P(HEMA MAAP-Ce3+)] cryogel was prepared by free radical polymerization method based on 2-hydroxyethyl methacrylic acid (HEMA) and methacryloyl antipyrin-Ce3+(MAAP Ce3+) complex. The MAAP-Ce3+ complex was characterized by UV-near infrared (UV-NIR) and energy dispersive X-ray (EDX) techniques, and the p(HEMA-MAAP Ce3+) cryogel was characterized by scanning electron microscopy (SEM), FT-IR and swelling tests. Five different peaks were seen at 916, 1037, 1081, 1109 and 1141 nm for the MAAP-Ce3+ complex in the UV-NIR spectrum. In the EDX spectrum, the presence of 15.12% N and 2.89% Ce3+ in the MAAP-Ce3+ complex showed nitrogen and Ce3+ in the structure. In FT-IR spectrum studies, cryogel showed –OH vibrations at 3412 cm-1 and characteristic –NH vibrations at 3104 cm 1 , and also at 1768-1712 cm-1 , ester, amide and cyclic carbonyl groups appears to have C=O bonds. The cryogel was used for lysozyme adsorption from the aqueous solution. Equilibrium swelling ratio of P(HEMA-MAAP-Ce3+) cryogel was calculated as 7.39 g H2O/g cryogel, and the macropore amount was calculated as 80.8%. The cryogel pore size was found to be 30-50 µm. The effects of the flow rate, pH, temperature, initial enzyme concentration and ionic strength on lysozyme adsorption were studied. Maximum adsorption capacity reached with 57.84 mg/g of prepared cryogel under 0,5 flow rate (mL/min), at pH 6.0 and at 25 oC. The result of the effect of ionic strength, it was determined that the lysozyme adsorption capacity decreased from 41.31 mg/g to 2.01 mg/g as the salt concentration increased from 0.05 M to 1.0 M. The correlation coefficient (R2 : 0.9939) obtained as a result of the values calculated for the Langmuir and Freundlich isotherms showed that the Langmuir adsorption model, could be suitable for this system. The adsorbed enzyme was desorbed 95.7% with 1.0 M NaCl solution containing 20 mM, pH 6.0 phosphate buffer in 120 minutes. In the prepared cryogel reusability studies, no significant decrease was observed in the adsorption capacity of the cryogel used 7 times in succession.
n this study, 2-hydroxyethyl methacrylic acid-methacryloyl antipyrin-Ce3+[P(HEMA MAAP-Ce3+)] cryogel was prepared by free radical polymerization method based on 2-hydroxyethyl methacrylic acid (HEMA) and methacryloyl antipyrin-Ce3+(MAAP Ce3+) complex. The MAAP-Ce3+ complex was characterized by UV-near infrared (UV-NIR) and energy dispersive X-ray (EDX) techniques, and the p(HEMA-MAAP Ce3+) cryogel was characterized by scanning electron microscopy (SEM), FT-IR and swelling tests. Five different peaks were seen at 916, 1037, 1081, 1109 and 1141 nm for the MAAP-Ce3+ complex in the UV-NIR spectrum. In the EDX spectrum, the presence of 15.12% N and 2.89% Ce3+ in the MAAP-Ce3+ complex showed nitrogen and Ce3+ in the structure. In FT-IR spectrum studies, cryogel showed –OH vibrations at 3412 cm-1 and characteristic –NH vibrations at 3104 cm 1 , and also at 1768-1712 cm-1 , ester, amide and cyclic carbonyl groups appears to have C=O bonds. The cryogel was used for lysozyme adsorption from the aqueous solution. Equilibrium swelling ratio of P(HEMA-MAAP-Ce3+) cryogel was calculated as 7.39 g H2O/g cryogel, and the macropore amount was calculated as 80.8%. The cryogel pore size was found to be 30-50 µm. The effects of the flow rate, pH, temperature, initial enzyme concentration and ionic strength on lysozyme adsorption were studied. Maximum adsorption capacity reached with 57.84 mg/g of prepared cryogel under 0,5 flow rate (mL/min), at pH 6.0 and at 25 oC. The result of the effect of ionic strength, it was determined that the lysozyme adsorption capacity decreased from 41.31 mg/g to 2.01 mg/g as the salt concentration increased from 0.05 M to 1.0 M. The correlation coefficient (R2 : 0.9939) obtained as a result of the values calculated for the Langmuir and Freundlich isotherms showed that the Langmuir adsorption model, could be suitable for this system. The adsorbed enzyme was desorbed 95.7% with 1.0 M NaCl solution containing 20 mM, pH 6.0 phosphate buffer in 120 minutes. In the prepared cryogel reusability studies, no significant decrease was observed in the adsorption capacity of the cryogel used 7 times in succession.
Açıklama
Anahtar Kelimeler
Kriyojel, İMAK, Seryum+3, Adsorpsiyon, Lizozim, Cryogel, IMAC, Cerium3+, Adsorption, Lysozyme
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Aksoy, E. (2023). Lizozim adsorpsiyonuna yönelik yeni bir kriyojelin hazırlanması ve enzim için bazı optimizasyon parametrelerinin belirlenmesi. Yayımlanmamış yüksek lisans tezi, Dicle Üniversitesi, Diyarbakır.
