Fe3O4@SiO2@NiAl-LDH microspheres implication in separation, kinetic and structural properties of phenylalanine dehydrogenase
Fe3O4@SiO2@NiAl-LDH three-components microsphere contains a Fe3O4@SiO2 magnetic core and a layered double hydroxide with nickel cation provide the binding ability to (His)-tagged-protein and exhibits high performance in protein separation and purification. The morphology and chemistry of the synthes...
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| Published in: | Heliyon Vol. 9; no. 9; p. e19429 |
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| Main Authors: | , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
Elsevier Ltd
01-09-2023
Elsevier |
| Subjects: | |
| Online Access: | Get full text |
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| Summary: | Fe3O4@SiO2@NiAl-LDH three-components microsphere contains a Fe3O4@SiO2 magnetic core and a layered double hydroxide with nickel cation provide the binding ability to (His)-tagged-protein and exhibits high performance in protein separation and purification. The morphology and chemistry of the synthesized Fe3O4@SiO2@NiAl-LDH microspheres were characterized by energy-dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), vibrating sample magnetometer (VSM), Dynamic light scattering (DLS). Purified enzyme was assesed with SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis and intrinsic fluorescence spectroscopy. In this study, the separation of phenylalanine dehydrogenase (PheDH) by Fe3O4@SiO2@NiAl -LDH was performed and the effect of microsphere was investigated on the kinetic and structural properties of PheDH. After purification, kinetic parameters such as Km, Vmax, Kcat, kcat/Km, optimum temperature, thermal stability, and and activation energy were evaluated and compared according to the mentioned methods. The interaction between the enzyme and the microsphere displayed a high performance in protein binding capacity. The results also revealed that the kinetic parameters of the enzyme changed in a dose-dependent manner in the presence of a microsphere. Moreover, the results of intrinsic fluorescence and Circular Dichroism (CD) confirmed the structural changes of the protein in the interaction with the microsphere.
In this work, we have exhibited high binding capacity of the phenylalanin dehydrogenase protein with Fe3O4@SiO2@NiAl-LDH microsphere, also we have compared kinetic parameters of phenylalanine dehydrogenase in presence of Fe3O4@SiO2@NiAl-LDH microsphere such as Kcat, kcat/km, km, Vm, thermal stability, optimum temperature and activation energy by utilizing the spectroscopic method. Also, it has been important to phenylalanine dehydrogenase protein purification via a simple process and using magnetic nanomaterials some advantages such as being readily available, and inexpensive. [Display omitted]
•The Protein separation is demonstrated using the high protein binding ability with (Fe3O4@SiO2@NiAl-LDH) magnetic nanoparticles.•The kinetic and thermochemical parameters of phenylalanine dehydrogenase are compared in presence of Fe3O4@SiO2@NiAl-LDH microsphere•Fe3O4@SiO2@NiAl-LDH microsphere can be used to separate His-tagged proteins. |
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| Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
| ISSN: | 2405-8440 2405-8440 |
| DOI: | 10.1016/j.heliyon.2023.e19429 |