Suda çözünebilen boronik asit fonksiyonel manyetik nanopartiküllerin sentezlenmesi ve L-asparajinaz immobilizasyonunda kullanımı
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2021
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info:eu-repo/semantics/openAccess
Özet
L-asparajinaz, başta Akut Lenfoblastik Lösemi olmak üzere birçok lenfoma hastalığının tedavisinde kulllanılan kemoterapötik bir enzim-ilaçtır. Bu tedavilerde kullanılan L-asparajinaz'ın bakterilerden izole edilmesinden dolayı kullanımını sınırlayan bazı yan etkileri mevcuttur. Bununlu birlikte L-asparajinazın hem kandaki yarı ömrünü arttırmak hem de daha etkili bir şekilde kullanılmasını sağlamak için çeşitli nanotaşıyıcı sistemler geliştirilmektedir. Bu çalışmanın amacı L-asparajinazı bor türevli manyetik nanopartiküllere immobilize etmek, immobilize enzimin optimum parametrelerini araştırmak ve zayıf manyetik alan altında indükleyerek enzimin katalitik aktivitesinin iyileştiğini göstermektir. Bu amaçla öncelikle taşıyıcı matriks olarak kullanılmak üzere manyetik demir oksit nanopartikülleri sentezlenmiş ve boronik asit ile modifiye edilmiştir. Çalışma kapsamında elde edilen tüm malzemeler yapısal ve morfolojik olarak karakterize edilmiştir. Ayrıca, immobilize enzimin optimum parametreleri (pH, sıcaklık), termal kararlılığı, depolama kararlılığı, yeniden kullanılabilirliği ve kinetik parametreleri incelenmiş ve serbest enzimle kıyaslanmıştır. Dahası, immobilize enzimin zayıf manyetik alan altında aktivitesi incelenmiştir. Serbest enzimin optimum pH ve sıcaklığı sırasıyla 8,5 ve 45 ℃ bulunurken, immobilize enzim için 7,5 ve 40 ℃ olarak bulunmuştur. Ayrıca, 10 kullanım döngüsünden sonra bile immobilize enzim başlangıç aktivitesinin %54'ünü korumuştur. Kinetik çalışmalarda serbest enzim için ve immobilize enzim içim Km değerleri sırasıyla 10,376±0,5 ve 7,06±2,99 mM olarak, Vmax ise sırasıyla 138,88±2,64 ve 121,95±1,070 µmol/dk olarak bulunmuştur. Bunlara ek olarak 10 Hz'lik bir manyetik alan ve 20 mT'lık bir manyetik akı uygulandığında aktivitenin sırasıyla yaklaşık 3,2 ve 4,3 kat arttığı gözlenmiştir. Sonuç olarak, L-asparajinaz immobilizasyonunda kullanılmak üzere suda çözünürlükleri yüksek fonksiyonel bor türevli manyetik nanotaşıyıcılar hazırlanmış, L-asparajinaz enzimi immobilize edilmiş ve immobilize enzimin aktivitesi zayıf manyetik alanda arttırılmıştır. Anahtar Kelimeler: Boronik asit, L-asparajinaz, Enzim immobilizasyonu, Nanotaşıyıcılar, Manyetik nanopartiküller
L-asparaginase is an enzyme-drug used in the treatment of Acute Lymphoblastic Leukemia (ALL) and some lymphomas. L-asparaginase, which is used in these treatments, has some side effects resulting from its isolation from bacteria and limiting its use. To prevent this situation, L-asparaginases are available in PEG-conjugated form. However, various magnetic nanocarrier systems are being developed to make L-asparaginase more biocompatible, to ensure that it reaches the desired region more effectively, and to examine its kinetic properties. The aim of this study is to synthesize boron-derived magnetic nanoparticles to immobilize L-asparaginase and to examine the kinetic properties of the immobilized enzyme. For this purpose, magnetic iron oxide nanoparticles were synthesized and modified with boronic acid to be used as carrier matrix. All materials obtained within the scope of the study were characterized structurally and morphologically. In addition, the optimum parameters (pH, temperature), thermal stability, storage stability, reusability and kinetic parameters of the immobilized enzyme were investigated and compared with the free enzyme. Moreover, the activity of the immobilized enzyme under weak magnetic field was investigated. The optimum pH and temperature of the free enzyme were found to be 8.5 and 45 °C, respectively, while it was found to be 7.5 and 40 °C for the immobilized enzyme. Moreover, even after 10 cycles of use, the immobilized enzyme retained 54% of its initial activity. In kinetic studies, Km values for free enzyme and for immobilized enzyme were found to be 10.376±0.5 and 7.06±2.99 mM, respectively, and Vmax was found to be 138.88±2.64 and 121.95±1.070 µmol/min, respectively. In addition, when a magnetic field of 10 Hz and a magnetic flux of 20 mT were applied, it was observed that the activity increased approximately 3,2 and 4,3 times, respectively. As a result, functional boron-derived magnetic nanocarriers with high water solubility were prepared for use in L-asparaginase immobilization, L-asparaginase enzyme was immobilized and the activity of the immobilized enzyme was increased in a weak magnetic field. Keywords: Boronic acid, L-asparaginase, Enzyme Immobilization, Nanocarriers, Magnetic nanoparticle
L-asparaginase is an enzyme-drug used in the treatment of Acute Lymphoblastic Leukemia (ALL) and some lymphomas. L-asparaginase, which is used in these treatments, has some side effects resulting from its isolation from bacteria and limiting its use. To prevent this situation, L-asparaginases are available in PEG-conjugated form. However, various magnetic nanocarrier systems are being developed to make L-asparaginase more biocompatible, to ensure that it reaches the desired region more effectively, and to examine its kinetic properties. The aim of this study is to synthesize boron-derived magnetic nanoparticles to immobilize L-asparaginase and to examine the kinetic properties of the immobilized enzyme. For this purpose, magnetic iron oxide nanoparticles were synthesized and modified with boronic acid to be used as carrier matrix. All materials obtained within the scope of the study were characterized structurally and morphologically. In addition, the optimum parameters (pH, temperature), thermal stability, storage stability, reusability and kinetic parameters of the immobilized enzyme were investigated and compared with the free enzyme. Moreover, the activity of the immobilized enzyme under weak magnetic field was investigated. The optimum pH and temperature of the free enzyme were found to be 8.5 and 45 °C, respectively, while it was found to be 7.5 and 40 °C for the immobilized enzyme. Moreover, even after 10 cycles of use, the immobilized enzyme retained 54% of its initial activity. In kinetic studies, Km values for free enzyme and for immobilized enzyme were found to be 10.376±0.5 and 7.06±2.99 mM, respectively, and Vmax was found to be 138.88±2.64 and 121.95±1.070 µmol/min, respectively. In addition, when a magnetic field of 10 Hz and a magnetic flux of 20 mT were applied, it was observed that the activity increased approximately 3,2 and 4,3 times, respectively. As a result, functional boron-derived magnetic nanocarriers with high water solubility were prepared for use in L-asparaginase immobilization, L-asparaginase enzyme was immobilized and the activity of the immobilized enzyme was increased in a weak magnetic field. Keywords: Boronic acid, L-asparaginase, Enzyme Immobilization, Nanocarriers, Magnetic nanoparticle
Açıklama
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İnönü Üniversitesi Fen Bilimleri Enstitüsü
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Künye
DİK, G. (2021). Suda çözünebilen boronik asit fonksiyonel manyetik nanopartiküllerin sentezlenmesi ve L-asparajinaz immobilizasyonunda kullanımı.
İnönü Üniversitesi, Yayınlanmış Yüksek Lisans Tezi.
