| Project/Area Number |
14350431
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | National University Corporation Tokyo University of Agriculture and Technology |
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Principal Investigator |
SODE Koji National University Corporation Tokyo University of Agriculture and Technology, Institute of Symbiotic Science and Technology, Professor, 大学院・共生科学技術研究部, 教授 (10187883)
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| Co-Investigator(Kenkyū-buntansha) |
IKEBUKURO Kazunori National University Corporation Tokyo University of Agriculture and Technology, Institute of Symbiotic Science and Technology, Associate Professor, 大学院・共生科学技術研究部, 助教授 (70251494)
FERRI Stefano National University Corporation Tokyo University of Agriculture and Technology, Institute of Symbiotic Science and Technology, Assistant Professor, 大学院・共生科学技術研究部, 助手 (90334474)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥14,200,000 (Direct Cost: ¥14,200,000)
Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2003: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2002: ¥7,000,000 (Direct Cost: ¥7,000,000)
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| Keywords | Enzyme reaction / Catalyst / chemical processes / Cytochrome / Nanobio / Electron transfer / Bioelectronics / Biosensors |
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| Research Abstract |
In most of oxidoreductases, the prosthetic group is buried deeply and within the protein shell. Thus a direct electrochemical recycling of the enzyme's prosthetic group at the electrode surface leading to a corresponding current signal is rarely encountered. In order to improve enzyme sensor signal property, we focused on the application of electron transfer proteins, cytochromes as the molecular interface between oxidorcductase redox center and electrode surface. In nature, various dehydrogenases harboring either PQQ or FAD are coupled with the respiratory chain via cytochromes. Based on this electron transfer property, we considered the utilization of cytochrome as the interface molecule to facilitate the electron transfer between enzyme and the artificial electron mediator or the electrode in the enzyme based biosensor. In this project, we focused on the application and engineering of cytochrome b_<562> as the interface molecule for novel bio-electronic devices and processes employi
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ng various oxidoreductases. The impact of the presence of cytochrome b_<562> on the various oxidoreductases was investigated, and its properties as the interface molecule were confirmed, by observing improved electrochemical response in the sensors employing these combinations. The engineering of cytochrome b_<562> was then attempted. The improvement of affinity toward PQQ glucose dehydrogenase was achieved by either modifying its surface charge or by introducing novel bio-affinity ligands. The employment of engineered interface molecules resulted in the bioelectrochemical reaction with PQQGDH, consequently improved the sensor properties. Finally, the co-expression of PQQ glucose dehydrogenase and cytochromeb_<562> was attempted to construct novel process employing interface molecules. The co-expression of PQQ glucose dehydrogenase and cytochromeb_<562> resulted in the construction of engineered E.coli cell which was able to transfer electron directly with electrode in the absence of artificial synthetic electron mediators. The results suggested the construction of novel bio-electrochemical process employing in vivo expressing interface molecules. Less
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