Comparative investigation of two methods for Acetylcholinesterase enzyme immobilization on modified porous silicon
| Autor: | Noure-Eddine Gabouze, S. Sam, Chafiaa Yaddaden, Amel Lounas, Khadidja Khaldi |
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| Rok vydání: | 2017 |
| Předmět: |
chemistry.chemical_classification
Immobilized enzyme Hydrosilylation Carboxylic acid General Physics and Astronomy 02 engineering and technology Surfaces and Interfaces General Chemistry 010402 general chemistry 021001 nanoscience & nanotechnology Condensed Matter Physics 01 natural sciences 0104 chemical sciences Surfaces Coatings and Films chemistry.chemical_compound Aminolysis chemistry Covalent bond Silanization Polymer chemistry Organic chemistry Peptide bond 0210 nano-technology Piranha solution |
| Zdroj: | Applied Surface Science. 421:148-154 |
| ISSN: | 0169-4332 |
| Popis: | In this work, Acetylcholinesterase enzyme (AChE) was immobilized on porous silicon (PSi) surface using two strategies. In the first method, acid chains were covalently grafted on the hydrogenated PSi by hydrosilylation reaction. The obtained acid-terminated surface was activated by a reaction with N -hydroxysuccinimide (NHS) in the presence of a peptide-coupling agent N -ethyl- N ′-(3-dimethylaminopropyl)-carbodiimide (EDC), and then reacted with the amino linker of the lysine residues AChE to anchor the enzyme by a covalent amide bond. In the second procedure, the PSi surface was first hydroxylated in piranha solution, followed by a silanization reaction with 3-aminopropyltriethoxysilane (APTES) to form amine-terminated surface. Finally, AChE was attached to the terminal amine groups by an aminolysis reaction with carboxylic acid groups of AChE in the presence of NHS/EDC mixture. Fourier transform infrared spectroscopy (FTIR) confirmed the efficiency of the surface modifications. The enzymatic activity of immobilized AChE was determined by means of a colorimetric test and was discussed according to the enzyme orientation on the surface which was revealed by contact angle measurements. |
| Databáze: | OpenAIRE |
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