Enzymes and proteins from extremophiles as hyperstable probes in nanotechnology: the use of D-trehalose/D-maltose-binding protein from the hyperthermophilic archaeon Thermococcus litoralis for sugars monitoring.
The D-trehalose/D-maltose-binding protein (TMBP), a monomeric protein of 48 kDa, is one component of the trehalose and maltose (Mal) uptake system. In the hyperthermophilic archaeon Thermococcus litoralis, this is mediated by a protein-dependent ATP-binding cassette system transporter. The gene coding for a thermostable TMBP from the archaeon T. litoralis has been cloned, and the recombinant protein has been expressed in E. coli. The recombinant TMBP has been purified to homogeneity and characterized. It exhibits the same functional and structural properties as the native one. In fact, it is highly thermostable and binds sugars, such as maltose, trehalose and glucose, with high affinity. In this work, we have immobilized TMBP on a porous silicon wafer. The immobilization of TMBP to the chip was monitored by reflectivity and Fourier Transformed Infrared spectroscopy. Furthermore, we have tested the optical response of the protein-Chip complex to glucose binding. The obtained data suggest the use of this protein for the design of advanced optical non-consuming analyte biosensors for glucose detection.
De Stefano
Luca
L
Istituto di Microelettrica e Microsistemi, CNR, Napoli, Italy.
Vitale
Annalisa
A
Rea
Ilaria
I
Staiano
Maria
M
Rotiroti
Lucia
L
Labella
Tullio
T
Rendina
Ivo
I
Aurilia
Vincenzo
V
Rossi
Mose'
M
D'Auria
Sabato
S
D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics; D015374:Biosensing Techniques / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000235:genetics; D005947:Glucose / Q000032:analysis*; D037102:Lectins / Q000737:chemistry* / Q000235:genetics; D008320:Maltose / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods; D016062:Porosity; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012825:Silicon / Q000737:chemistry; D019712:Thermococcus / Q000201:enzymology* / Q000235:genetics; D014199:Trehalose / Q000737:chemistry
D019843:Archaeal Proteins; D004800:Enzymes, Immobilized; D037102:Lectins; D011994:Recombinant Proteins; D008320:Maltose; D014199:Trehalose; D005947:Glucose; D012825:Silicon