超好熱菌酵素を素子とするバイオセンサーの開発 : ポリアミン関連酵素の機能解析とD-プロリン脱水素酵素機能電極センサーの開発 [in Japanese] Development of Biosensors Using Hyperthermophilic Enzymes as an Element:Development of Biosensors Using Hyperthermophilic Enzymes as an Element:Functional Analysis of Polyamine Metabolizing Enzymes and Development for EnzymeElectrode Sensor Using D-Proline Dehydrogenases [in Japanese]
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An amperometric enzyme sensor give us higher sensitivity and specificity for the substratedetermination. In spite of advantages of enzyme sensor, many enzymes so far found have been toolabile to use as biosensor elements in artificial circumstances for a longer period.Hyperthermophiles, which can grow above 90℃, have been known to produce much more stableenzymes under various artificial conditions. In this work, we carried out screening, biochemicalcharacterization and improvement of production for hyperthermostable enzymes which are more usefulas the elements in the biosensors. We focused on the polyamines as one of the substrates ofbiosensors. Polyamines have been known to play many important roles in cell proliferation,differentiation and transformation. The concentration of the polyamines together with their acetylconjugates remarkably increases in the biological fluids and the affected tissues of cancer patients.Therefore, their polyamines are listed as tumor markers. Gas and ion chromatographies have beenso far used for polyamine determination, but have some problems from the aspects of high sensitivityand easy operation. Thus, we here developed biosensors using hyperthermostable enzymes forpolyamine determination. Such enzyme sensors are more useful for the simple and rapiddetermination of polyamines and application for clinical analysis and food analysis In addition, wetried the construction of biosensor using the hyperthermophilic enzyme, D-Proline dehydrogenase.As the results, we found the thermostable agmatinase and spermidine dehydrogenase inhyperthermophiles, Pyrococcus horikoshii and Sulfolobus tokodaii, respectively. We succeeded theconstruction of novel amperometric sensor for D-proline determination using D-Proline dehydrogenasederived from Pyrobaculum islandicum.
- Bulletin of Faculty of Engineering the University of Tokushima
Bulletin of Faculty of Engineering the University of Tokushima (50), 2005-03
The University of Tokushima