http://www.cnr.it/ontology/cnr/individuo/prodotto/ID172732
A Thermostable Sugar-Binding Protein from the Archaeon Pyrococcus horikoshii as a Probe for the Development of a Stable Fluorescence Biosensor for Diabetic Patients (Articolo in rivista)
- Type
- Label
- A Thermostable Sugar-Binding Protein from the Archaeon Pyrococcus horikoshii as a Probe for the Development of a Stable Fluorescence Biosensor for Diabetic Patients (Articolo in rivista) (literal)
- Anno
- 2004-01-01T00:00:00+01:00 (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi
- 10.1021/bp049857g (literal)
- Alternative label
Staiano M.; Sapio M.; Scognamiglio V.; Marabotti A.; Facchiano A.M.; Bazzicalupo P.; Rossi M.; D'Auria S. (2004)
A Thermostable Sugar-Binding Protein from the Archaeon Pyrococcus horikoshii as a Probe for the Development of a Stable Fluorescence Biosensor for Diabetic Patients
in Biotechnology progress (Print); ACS, American chemical society, Washington, DC (Stati Uniti d'America)
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- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
- Staiano M.; Sapio M.; Scognamiglio V.; Marabotti A.; Facchiano A.M.; Bazzicalupo P.; Rossi M.; D'Auria S. (literal)
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- Rivista
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo
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- ISI Web of Science (WOS) (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
- Institute of Protein Biochemistry, CNR Via Pietro Castellino, 111 80131 Naples, Italy,
Institute of Genetics & Biophysics, CNR, Naples, Italy,
Laboratory of Bioinformatics, Institute of Food Science, CNR, Avellino, Italy, and Centro di Ricerca Interdipartimentale di Scienze Computazionali E Biotecnologiche (CRISCEB),
Seconda Universita` di Napoli, Naples, Italy, and Center for Fluorescence Spectroscopy, UMAB, Baltimore, Maryland (literal)
- Titolo
- A Thermostable Sugar-Binding Protein from the Archaeon Pyrococcus horikoshii as a Probe for the Development of a Stable Fluorescence Biosensor for Diabetic Patients (literal)
- Abstract
- In this work is presented the first attempt to develop an innovative ultrastable protein-based biosensor for blood glucose detections. The gene of a putative thermostable sugar-binding protein has been cloned and expressed in E. coli. The recombinant protein has been purified to homogeneity by thermoprecipitation and affinity chromatography steps. The recombinant protein is a monomer with an apparent molecular weight of 55,000 as judged by gel filtration and sodium dodecyl sulfate polyacrylamide gel eletrophoresis. Circular dichroism experiments showed that the protein possesses a secondary structure content rich in alpha-helices and beta-structures and that the protein is highly stable as investigated in the range of temperature between 20 and 95 degrees C. Fluorescence spectroscopy experiments demonstrated that the recombinant protein binds glucose with a dissociation constant of about 10 mM, a concentration of sugar very close to the concentration of glucose present in the human blood. A docking simulation on the modeled structure of the protein confirms its ability to bind glucose and proposes possible modifications to improve the affinity for glucose and/or its detection. The obtained results suggest the use of the protein as a probe for a stable glucose biosensor. (literal)
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