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Examination of chondroitinase ABC I immobilization onto dextran-coated Fe3O4 nanoparticles and its in-vitro release

Askaripour, H ; Sharif University of Technology | 2020

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  1. Type of Document: Article
  2. DOI: 10.1016/j.jbiotec.2019.12.020
  3. Publisher: Elsevier B.V , 2020
  4. Abstract:
  5. Chondroitinase ABC I (cABC I) has received notable attention in treatment of spinal cord injuries and its application as therapeutics has been limited due to low thermal stability at physiological temperature. In this study, cABC I enzyme was immobilized on the dextran-coated Fe3O4 nanoparticles through physical adsorption to improve the thermal stability. The nanoparticles were characterized using XRD, SEM, VSM, and FTIR analyses. Response surface methodology and central composite design were employed to assess factors affecting the activity of immobilized cABC I. Experimental results showed that pH 6.3, temperature 24 °C, enzyme/support mass ratio 1.27, and incubation time 5.7 h were the optimal immobilization conditions. It was found that thermal stability of immobilized cABC I was significantly improved. In-vitro cABC I release was studied under pH 7.5 and temperature 37 °C and the results indicated that 70 % release occurred after 9 h and the release mechanism was first-order kinetic model. © 2020 Elsevier B.V
  6. Keywords:
  7. Chondroitinase ABC I enzyme ; Dextran-coated Fe3O4nanoparticles ; In-vitrorelease ; Release mechanism ; Response surface methodology ; Dextran ; Iron oxides ; Magnetite ; Nanoparticles ; Surface properties ; Thermodynamic stability ; Central composite designs ; Chondroitinase abc's ; First-order kinetic models ; Immobilization conditions ; Physiological temperature ; Spinal cord injuries (SCI) ; Enzyme immobilization ; Chondroitin ABC lyase 1 ; Unclassified drug ; Immobilized enzyme ; Magnetite nanoparticle ; Article ; Conformational transition ; Enzyme purification ; Fourier transform infrared spectroscopy ; In vitro study ; Magnetic field ; Michaelis Menten kinetics ; Nonhuman ; Particle size ; PH ; Polyacrylamide gel electrophoresis ; Priority journal ; Protein expression ; Response surface method ; Scanning electron microscopy ; Substrate concentration ; Surface property ; Synthesis ; Thermostability ; Vibrating sample magnetometry ; X ray diffraction ; Chemistry ; Genetics ; Metabolism ; Adsorption ; Chondroitin ABC Lyase ; Dextrans ; Enzyme Stability ; Enzymes, Immobilized ; Escherichia coli ; Hydrogen-Ion Concentration ; Kinetics ; Magnetite Nanoparticles ; Proteus vulgaris ; Temperature ; X-Ray Diffraction
  8. Source: Journal of Biotechnology ; Volume 309 , 2020 , Pages 131-141
  9. URL: https://www.sciencedirect.com/science/article/abs/pii/S0168165619309642