Üreazın modifiye edilmiş florisile kovalent immobilizasyonu ve serbest ve immobilize üreazın karakterizasyonu

Özlem Alptekin

doi:10.17826/cumj.453980

TR EN

Üreazın modifiye edilmiş florisile kovalent immobilizasyonu ve serbest ve immobilize üreazın karakterizasyonu

Öz

Amaç:Bu çalışmada Canavalia ensiformis kaynaklı üreazın modifiye edilmiş Florisil’e (magnezyum silikat) kovalent immobilize edilmesi ve immobilize üreazın karakterizasyonunun yapılarak biyoreaktör uygulamalarında kullanılabilirliğinin araştırılması amaçlanmıştır.

Gereç ve Yöntem:Florisil, 3-aminopropiltrietoksisilan ile aktifleştirilmiş daha sonra glutaraldehit ile modifiye edilmiştir. Modifiye edilmiş desteğe üreaz kovalent immobilize edilmiştir. İmmobilize enzimin ve desteğin taramalı elektron mikroskobunda görüntüleri incelenmiştir. Serbest ve immobilize üreazın optimum pH’sı, sıcaklığı, kinetik parametreleri (Km, Vmax, kcat/Km) belirlenmiş, 4°C’de ve oda sıcaklığında depolama kararlılıkları incelenmiştir. İmmobilize üreazın kesikli tip reaktörde tekrar kullanılabilirliği araştırılmıştır.

Bulgular:Serbest ve immobilize üreaz için optimum pH sırasıyla 7,0 ve 6,5; sıcaklık ise 50 ve 60°C olarak belirlenmiştir. İmmobilizasyondan sonra enzimin Km değeri 2,2 kat artmıştır. İmmobilize üreazın katalitik etkinliği, serbest üreazın katalitik etkinliğinin %.0,1’i kadar bulunmuştur. Oda sıcaklığında ve 4°C’de serbest üreaz 5 gün sonunda ve aktivitesini tamamen kaybederken immobilize üreaz 19 gün sonunda aktivitesini kaybetmemiştir. İmmobilize üreaz kesikli tip biyoreaktörde 10 kullanım sonunda başlangıç aktivitesinin %50’sini korumuştur.

Sonuç:Modifiye Florisile kovalent immobilize edilen üreazın serbest üreaza göre düşük katalitik aktivite göstermesine rağmen, immobilize üreazın serbest üreaza göre depolama kararlılığının yüksek olması ve biyoreaktör uygulamalarında tekrar kullanılabilmesi nedeniyle immobilize üreazın kullanım potansiyeli yüksektir.

Anahtar Kelimeler

Üreaz,immobilizasyon,bioreaktör

Covalent immobilization of urease onto modified florisil and characterization of free and immobilized urease

Abstract

Purpose: In this study, it was aimed to immobilize urease from Canavalia ensiformis onto modified Florisil (magnesium silicate) by covalently and to characterize immobilized urease and also to investigate reusability of immobilized urease in a batch type bioreactor.

Materials and Methods: Florisil was activated with 3-aminopropyltriethoxysilane and then modified with glutaraldehyde. Urease was covalently immobilized onto modified support. Images of support and immobilized urease were examined on a scanning electron microscope. The optimum pH and temperature, kinetic parameters (Km, Vmax, kcat/Km) and storage stabilities at 4°C and room temperature of free and immobilized ureases were determined. Reusability of immobilized urease was investigated in a batch type bioreactor.

Results: Free and immobilized ureases showed their maximum activities at pHs 7.0 and 6.5, and at temperatures 50 and 60°C, respectively. The Km value of immobilized urease increased 2.2 fold compared to free urease. The catalytic efficiency of immobilized urease was about 0.1% of the free urease. Free urease completely lost its activity at the end of 5 days stored at 4°C and room temperature; however, immobilized urease did not lose its activity at the end of 19 days at the same conditions. Immobilized urease maintained 50% of its initial activity at the end of 10 reuses in batch type bioreactor.

Conclusion: Although urease activity significantly decreased upon covalent immobilization onto modified Florisil. However, high storage stability and reusability of immobilized urease makes it potentially useful immobilized form.

Keywords

Urease,immobilization,bioreactor,Florisil

Kaynakça

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Ayrıntılar

Birincil Dil

Türkçe

Konular

Sağlık Kurumları Yönetimi

Bölüm

Araştırma Makalesi

Yazarlar

Özlem Alptekin ^*
0000-0002-0458-7609
Türkiye

Yayımlanma Tarihi

30 Eylül 2019

Gönderilme Tarihi

16 Ağustos 2018

Kabul Tarihi

6 Ocak 2019

Yayımlandığı Sayı

Yıl 2019 Cilt: 44 Sayı: 3

DOI

https://doi.org/10.17826/cumj.453980

IZ

https://izlik.org/JA46PT56PR

MLA

Alptekin, Özlem. “Üreazın modifiye edilmiş florisile kovalent immobilizasyonu ve serbest ve immobilize üreazın karakterizasyonu”. Cukurova Medical Journal, c. 44, sy 3, Eylül 2019, ss. 811-8, doi:10.17826/cumj.453980.