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.
De Stefano, Luca ; Vitale, Annalisa ; Rea, Ilaria ; et al. ; - ASI Sponsor
Jan - 2008
ISSN : 1431-0651 ;
journal : Extremophiles : life under extreme conditions

Issue : 1
type: Article Journal

Abstract
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.

keywords : Archaeal Proteins,Archaeal Proteins: chemistry,Archaeal Proteins: genetics,Biosensing Techniques,Biosensing Techniques: methods,Enzymes,Glucose,Glucose: analysis,Immobilized,Immobilized: chemistry,Immobilized: genetics,Lectins,Lectins: chemistry,Lectins: genetics,Maltose,Maltose: chemistry,Nanotechnology,Nanotechnology: methods,Porosity,Recombinant Proteins,Recombinant Proteins: chemistry,Recombinant Proteins: genetics,Silicon,Silicon: chemistry,Thermococcus,Thermococcus: enzymology,Thermococcus: genetics,Trehalose,Trehalose: chemistry