Journals / Hacettepe Journal of Biology and Chemistry / 2019 / Cilt: 47 - Sayı: 1

Biologically Modified poly(2-hydroxylethyl methacrylate) Cryogels for Lysozyme Purification

Lizozim Saflaştırılması İçin Heparin İmmobilize Edilmiş poli(2-hidroksi etilmetakrilat) Kriyojeller

Pages
123–131
Publication Date
2022-10-17
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

The purity of lysozyme from egg white studies wasanalysed by sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using 12% separating gel.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Heparin immobilized poly(2-hydroxylethyl methacrylate) PHEMA cryogel was synthesized and applied for lysozymepurification from egg white.
📊
Key Findings & Data:Thesurface area of PHEMA cryogel was found to be 25.2 m2/g.
💡
Conclusion & Impact:The purity of lysozyme from egg white studies wasanalysed by sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using 12% separating gel.
🔬Design:Modeling & Simulation
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Heparin immobilized poly(2-hydroxylethyl methacrylate) PHEMA cryogel was synthesized and applied for lysozymepurification from egg white. Firstly, the PHEMA cryogel was synthesized by cryopolymerization and then heparin wascovalently immobilized on to the PHEMA cryogel with cyanogen bromide activation. The modification of PHEMA cryogelstructure with heparin was further confirmed by Fourier-transform infrared spectroscopy (FTIR). The surface and innerstructure morphologies of PHEMA cryogels were studied and characterized by the scanning electron microscope (SEM). Thesurface area of PHEMA cryogel was found to be 25.2 m2/g. Heparin immobilized PHEMA cryogels were used in lysozymeadsorption studies to assess the effects of pH, lysozyme concentration, flow rate, temperature and ionic strength. Themaximum lysozyme adsorption on the heparin immobilized PHEMA cryogel was found to be 48.73 mg/g from aqueoussolutions under optimized conditions. 1.0 M NaCI solution was used for desorption of lysozyme in a continuous system.The reusability of heparin immobilized PHEMA cryogels was tested for 10 adsorption-desorption cycles. The Langmuiradsorption model was plotted and found fitted for adsorption studies. The purity of lysozyme from egg white studies wasanalysed by sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using 12% separating gel.

Özet

Yumurta akından lizozim saflaştırılması için heparin immobilize edilmiş poli(hidroksi etilmetakrilat) (PHEMA) kriyojeller sentezlenmiştir. İlk olarak, PHEMA kriyojeller kriyopolimerizasyon yöntemi ile sentezlenmiştir. Heparin, siyanojen bromür aktivasyonu ile PHEMA kriyojeline kovalent olarak immobilize edilmiştir. PHEMA kriyojel yapısının heparin ile modifikasyonu Fourier dönüşümlü kızıl ötesi spektroskopisi (FTIR) ile incelenmiştir. PHEMA kriyojellerin yüzey ve iç yapı morfolojileri taramalı elektron mikroskobu (SEM) ile karakterize edilmiştir. PHEMA kriyojelin yüzey alanı 25.2 m2 /g olarak bulunmuştur. Heparin immobilize edilmiş PHEMA kriyojeller lizozim adsorpsiyon çalışmalarında pH, lizozim derişimi, akış hızı, sıcaklık ve iyonik şiddetin etkilerini incelemek için kullanılmıştır. Optimum koşullar altında, heparin immobilize edilmiş PHEMA kriyojelin sulu çözeltide en yüksek lizozim adsorpsiyonu 48.73 mg/g olarak bulunmuştur. Sürekli sistemde lizozimin desorpsiyonu için 1.0 M NaCI çözeltisi kullanılmıştır. Heparin immobilizasyonuna heparin derişiminin etkisi incelenmiştir. Heparin immobilize edilmiş PHEMA kriyojellerin tekrar kullanılabilirliği adsorpsiyon-desorpsiyon döngüsü 10 kez tekrarlanarak test edilmiştir. Langmuir adsorpsiyon modeli çizilmiş ve adsorpsiyon çalışmaları için uygun bulunmuştur. Yumurta akından lizozim saflığı %12 ayırıcı jel kullanılarak sodyum-dodesil sülfat poliakrilamid jel elektroforezi kullanılarak analiz edilmiştir.

🔖

Cite This Article

ÇİMEN, D., DENİZLİ, A. (2022). Biologically Modified poly(2-hydroxylethyl methacrylate) Cryogels for Lysozyme Purification. Hacettepe Journal of Biology and Chemistry, Cilt: 47 - Sayı: 1, 123–131. https://ajindex.com/articles/529010

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (14)

  1. M. Andac, I.Y. Galaev, A. Denizli, Affinity based and molecularly imprinted cryogels: Applications in biomacromolecule purification, J. Chromatogr. B, 1021 (2016) 69-80
  2. M. Uygun, D.A. Uygun, E. Özçalışkan, S. Akgöl, A. Denizli, Concanavalin A immobilized poly(ethylene glycol dimethacrylate) based affinity cryogel matrix and usability of invertase immobilization, J. Chromatogr. B, 73 (2012) 887- 888
  3. S. Hajizadeh, C. Xu, H. Kirsebom, L. Ye, B. Mattiasson, Cryogelation of molecularly imprinted nanoparticles: A macroporous structure as affinity chromatography column for removal of β-blockers from complex samples. J. Chromatogr. A, 1274 (2013) 6-12
  4. W. Noppe, H. Deckmyn, Development and screening of epoxy-spacer-phage cryogels for affinity chromatography: Enhancing the binding capacity, J. Sep. Sci., 40 (2017) 2575- 2583
  5. V.I., Lozinsky, Polymeric Cryogels as a new family of macroporous and supermacroporous materials for biotechnological purposes, Russ. Chem. Bull., 57 (2008) 1015-1032
  6. F.M. Plieva, H. Kirsebom, B. Mattiasson, Preparation of macroporous cryostructurated gel monoliths, their characterization and main applications, J. Sep. Sci., 34 (2011) 2164-2172
  7. H. Kirsebom, I.Y. Galaev, B. Mattiasson, Stimuli-responsive polymers in the 21st century: elaborated architecture to achieve high sensitivity, fast response, and robust behavior, J. Polym. Sci. Pol. Phys., 49 (2011) 173-178
  8. N. Bereli, M. Andaç, G. Baydemir, R. Say, I.Y. Galaev, A. Denizli, Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels, J. Chrom. A, 1190 (2008) 18-26
  9. M. Andaç, I.Y. Galaev, A. Denizli, Dye attached poly(hydroxyethyl methacrylate) cryogel for albumin depletion from human serum, J. Sep. Sci., 35 (2012) 1173- 1182
  10. V.M. Gun’ko, I.N. Savina, S.V. Mikhalovsky, Cryogels: morphological structural and adsorption characterization, Adv. Colloid Interface Sci., 187 (2013) 1-46
  11. H. Alkan, N. Bereli, Z. Baysal, A. Denizli, Antibody purification with protein A attached supermacroporous poly(hydroxyethyl methacrylate) cryogel, Biochem. Eng. J., 45 (2009) 201-208
  12. D. Çimen, A. Denizli, Immobilized metal affinity monolithic cryogels for cytochrome C purification, Colloid. Surf. B Biointer., 93 (2012) 29-35
  13. E. Tamahkar, N. Bereli, R. Say, A. Denizli, Molecularly imprinted supermacroporous cryogels for cytochrome C recognition, J. Sep. Sci., 34 (2011) 3433-3440
  14. N. Bereli, M. Andaç, G. Baydemir, R. Say, I.Y. Gala