Int. J. Mol. Sci. 2013, 14(3), 4613-4628; doi:10.3390/ijms14034613
Article

Covalent Immobilization of Bacillus licheniformis γ-Glutamyl Transpeptidase on Aldehyde-Functionalized Magnetic Nanoparticles

1,†email, 1,†email, 1email, 2,* email and 1,* email
Received: 15 January 2013; in revised form: 20 February 2013 / Accepted: 21 February 2013 / Published: 26 February 2013
(This article belongs to the Special Issue Magnetic Nanoparticles 2013)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: This work presents the synthesis and use of surface-modified iron oxide nanoparticles for the covalent immobilization of Bacillus licheniformis γ-glutamyl transpeptidase (BlGGT). Magnetic nanoparticles were prepared by an alkaline solution of divalent and trivalent iron ions, and they were subsequently treated with 3-aminopropyltriethoxysilane (APES) to obtain the aminosilane-coated nanoparticles. The functional group on the particle surface and the amino group of BlGGT was then cross-linked using glutaraldehyde as the coupling reagent. The loading capacity of the prepared nanoparticles for BlGGT was 34.2 mg/g support, corresponding to 52.4% recovery of the initial activity. Monographs of transmission electron microscopy revealed that the synthesized nanoparticles had a mean diameter of 15.1 ± 3.7 nm, and the covalent cross-linking of the enzyme did not significantly change their particle size. Fourier transform infrared spectroscopy confirmed the immobilization of BlGGT on the magnetic nanoparticles. The chemical and kinetic behaviors of immobilized BlGGT are mostly consistent with those of the free enzyme. The immobilized enzyme could be recycled ten times with 36.2% retention of the initial activity and had a comparable stability respective to free enzyme during the storage period of 30 days. Collectively, the straightforward synthesis of aldehyde-functionalized nanoparticles and the efficiency of enzyme immobilization offer wide perspectives for the practical use of surface-bound BlGGT.
Keywords: Bacillus licheniformis; γ-glutamyl transpeptidase; magnetic nanoparticle; 3-aminopropyltriethoxysilane; covalent immobilization
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MDPI and ACS Style

Chen, Y.-Y.; Tsai, M.-G.; Chi, M.-C.; Wang, T.-F.; Lin, L.-L. Covalent Immobilization of Bacillus licheniformis γ-Glutamyl Transpeptidase on Aldehyde-Functionalized Magnetic Nanoparticles. Int. J. Mol. Sci. 2013, 14, 4613-4628.

AMA Style

Chen Y-Y, Tsai M-G, Chi M-C, Wang T-F, Lin L-L. Covalent Immobilization of Bacillus licheniformis γ-Glutamyl Transpeptidase on Aldehyde-Functionalized Magnetic Nanoparticles. International Journal of Molecular Sciences. 2013; 14(3):4613-4628.

Chicago/Turabian Style

Chen, Yi-Yu; Tsai, Ming-Gen; Chi, Meng-Chun; Wang, Tzu-Fan; Lin, Long-Liu. 2013. "Covalent Immobilization of Bacillus licheniformis γ-Glutamyl Transpeptidase on Aldehyde-Functionalized Magnetic Nanoparticles." Int. J. Mol. Sci. 14, no. 3: 4613-4628.

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