Formation and characterization of glutamate dehydrogenase monolayers on silicon supports.
In this paper we have tested two different procedures (the "three-step" and the "four-step" procedures) for the covalent immobilization of glutamate dehydrogenase (GDH) onto silicon supports. Atomic force microscopy (AFM), Fourier-transform infrared spectroscopy (FT-IR), fluorescence spectroscopy and an enzymatic assay were used to probe the structure and activity of the immobilized enzyme. Our results demonstrate that coupling through the "three-step" procedure does not significantly affect either the fold pattern or the activity of the enzyme, suggesting that this method could be ideally suited to the development of high quality monolayers for use in enzyme-based planar biosensors.
Blasi
L
L
National Nanotechnology Laboratory of INFM, c/o Department of Innovation Engineering, University of Lecce, Italy. laura.blasi@unile.it
Longo
L
L
Pompa
P P
PP
Manna
L
L
Ciccarella
G
G
Vasapollo
G
G
Cingolani
R
R
Rinaldi
R
R
Rizzello
A
A
Acierno
R
R
Storelli
C
C
Maffia
M
M
D015374:Biosensing Techniques / Q000295:instrumentation*; D004800:Enzymes, Immobilized; D005969:Glutamate Dehydrogenase; D018625:Microscopy, Atomic Force; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D012825:Silicon; D017550:Spectroscopy, Fourier Transform Infrared
D004800:Enzymes, Immobilized; D009243:NAD; D005969:Glutamate Dehydrogenase; D012825:Silicon