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.
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Acknowledgments
The authors thank Dr. Fabienne Chevance, University of Utah, Salt Lake City, USA, and Prof. Winfried Boos, University of Konstanz, Germany, for supplying a partially purified protein sample. This work is in the frame of the project “Diagnostica Avanzata ed Alimentazione” CNR Commessa of the Agro-Food Department (SD). This work was also supported by the ASI project MoMa No. 1/014/06/0 (SD) and by a grant from the Ministero degli Affari Esteri, Direzione Generale per la Promozione e la Cooperazione Culturale (SD).
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Communicated by D. A. Cowan.
The authors wish to dedicate this work to Prof. Ignacy Gryczynski, University of North Texas, TX, USA, for his outstanding contribution to the development of new sensing methodologies.
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De Stefano, L., Vitale, A., Rea, I. et al. 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. Extremophiles 12, 69–73 (2008). https://doi.org/10.1007/s00792-006-0058-6
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DOI: https://doi.org/10.1007/s00792-006-0058-6