Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles
Over the last decade, nanoparticles used as protein carriers have opened new avenues for a variety of biomedical applications. The main concern for these applications is changes in biological activity of immobilized proteins due to conformational changes on the surface of the carrier. To evaluate th...
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| Format: | Article |
| Language: | English |
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Wiley
2013-01-01
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| Series: | Journal of Nanotechnology |
| Online Access: | http://dx.doi.org/10.1155/2013/830151 |
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| author | Mahmoud Kamal Ahmadi Manouchehr Vossoughi |
| author_facet | Mahmoud Kamal Ahmadi Manouchehr Vossoughi |
| author_sort | Mahmoud Kamal Ahmadi |
| collection | DOAJ |
| description | Over the last decade, nanoparticles used as protein carriers have opened new avenues for a variety of biomedical applications. The main concern for these applications is changes in biological activity of immobilized proteins due to conformational changes on the surface of the carrier. To evaluate this concern, the preparation and biocatalyst activity of α-chymotrypsin-Fe3O4 @ Au core/shell nanoparticles were investigated. First, Fe3O4 @ Au core/shell nanoparticles were synthesized by coprecipitation method and citrate reduction of HAuCl4. TEM imaging revealed a core size of 13 ± 3 nm and a shell thickness of 4 ± 1 nm for synthesized nanoparticles. X-ray diffraction (XRD) was used to study the crystalline structure of the nanoparticles. Next, the enzyme was immobilized on the surface of synthesized nanoparticles by covalent bonding of Au shell with thiol and amine groups present in the protein structure (e.g., cysteine and histidine residues). FTIR and fluorescence spectroscopy were utilized to study secondary and tertiary structures of the immobilized enzyme. Results show that the secondary and tertiary structures of the enzyme remain virtually unchanged after immobilization on the nanoparticles surface. However, the biocatalyst activity of the enzyme was reduced by thirty percent, indicating possible conformational changes or active site occlusion. |
| format | Article |
| id | doaj-art-b9aaf8931e314ca28ae755dd072eb26a |
| institution | Kabale University |
| issn | 1687-9503 1687-9511 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Nanotechnology |
| spelling | doaj-art-b9aaf8931e314ca28ae755dd072eb26a2025-02-03T01:01:14ZengWileyJournal of Nanotechnology1687-95031687-95112013-01-01201310.1155/2013/830151830151Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell NanoparticlesMahmoud Kamal Ahmadi0Manouchehr Vossoughi1Department of Chemical and Biological Engineering, State University of New York at Buffalo, 303 Furnas Hall, Buffalo, NY 14260-4200, New York, USADepartment of Chemical and Petroleum Engineering, Sharif University of Technology, Azadi Avenue, P.O. Box 11365, Tehran, IranOver the last decade, nanoparticles used as protein carriers have opened new avenues for a variety of biomedical applications. The main concern for these applications is changes in biological activity of immobilized proteins due to conformational changes on the surface of the carrier. To evaluate this concern, the preparation and biocatalyst activity of α-chymotrypsin-Fe3O4 @ Au core/shell nanoparticles were investigated. First, Fe3O4 @ Au core/shell nanoparticles were synthesized by coprecipitation method and citrate reduction of HAuCl4. TEM imaging revealed a core size of 13 ± 3 nm and a shell thickness of 4 ± 1 nm for synthesized nanoparticles. X-ray diffraction (XRD) was used to study the crystalline structure of the nanoparticles. Next, the enzyme was immobilized on the surface of synthesized nanoparticles by covalent bonding of Au shell with thiol and amine groups present in the protein structure (e.g., cysteine and histidine residues). FTIR and fluorescence spectroscopy were utilized to study secondary and tertiary structures of the immobilized enzyme. Results show that the secondary and tertiary structures of the enzyme remain virtually unchanged after immobilization on the nanoparticles surface. However, the biocatalyst activity of the enzyme was reduced by thirty percent, indicating possible conformational changes or active site occlusion.http://dx.doi.org/10.1155/2013/830151 |
| spellingShingle | Mahmoud Kamal Ahmadi Manouchehr Vossoughi Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles Journal of Nanotechnology |
| title | Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles |
| title_full | Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles |
| title_fullStr | Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles |
| title_full_unstemmed | Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles |
| title_short | Immobilization of α-Chymotrypsin on the Surface of Magnetic/Gold Core/Shell Nanoparticles |
| title_sort | immobilization of α chymotrypsin on the surface of magnetic gold core shell nanoparticles |
| url | http://dx.doi.org/10.1155/2013/830151 |
| work_keys_str_mv | AT mahmoudkamalahmadi immobilizationofachymotrypsinonthesurfaceofmagneticgoldcoreshellnanoparticles AT manouchehrvossoughi immobilizationofachymotrypsinonthesurfaceofmagneticgoldcoreshellnanoparticles |