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

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

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

Int. J. Mol. Sci. 2013, 14, 4613-4628; doi:10.3390/ijmOPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 /journal/ijms Article Covalent Immobilization of Bacillus licheniformis γ-Glutamyl Transpeptidase on Aldehyde-Functionalized Magnetic Nanoparticles Yi-Yu Chen 1,?, Ming-Gen Tsai 1,?, Meng-Chun Chi , Tzu-Fan Wang * and Long-Liu Lin 1,* 1 2, 1 Department of Applied Chemistry, National Chiayi University, 300 Syuefu Road, Chiayi City 60004, Taiwan; E-Mails: s0972236@.tw (Y.-Y.C.); s0992739@.tw (M.-G.T.); s0910324@.tw (M.-C.C.) 2 Department of Life Sciences and Institute of Molecular Biology, National Chung Cheng University, Chiayi County 621, Taiwan ? These authors contributed equally to this work. * Authors to whom correspondence should be addressed; E-Mails: catalpa0905@ (T.-F.W.); llin@.tw (L.-L.L.); Tel.: +886-5271-7969 (L.-L.L.); Fax: +886-5271-7901 (L.-L.L.). Received: 15 January 2013; in revised form: 20 February 2013 / Accepted: 21 February 2013 / Published: 26 February 2013 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