| Project/Area Number |
09680634
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | Kansai Medical Universtiy |
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Principal Investigator |
FUJII Shigeru Kansai Medical University, Professor, 医学部, 教授 (60144482)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAGAWA Manabu Kansai Medical University, Assistant, 医学部, 助手 (50261053)
KURODA Kiyo Kansai Medical University, Assistant, 医学部, 助手 (30131436)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Flavoenzyme / Reaction Mechanism / Electron Transfer / NMR / Stable Isotope Labeling / ワラビン酵素 |
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| Research Abstract |
The results of this research project were summarized as follows. It was confirmed that old yellow enzyme (OYE) exists in cytosol of brewer's yeast cell, In the fermentation process using the brewer's yeast, the amount of OYE rapidly increased during the first 12 hrs of the fermentation and then gradually decreased in a low oxygen concentration. These results suggest that OYE works in the presence of oxygen. Old Yellow enzyme (OYB) from brewer's yeast was reconstituted with ^<13>C-enriched FMNs and 8-fluoro-8-demethyl-FMN (8F-FMN). Interactions of the reconstituted OYEs with NADP^+ and with cytochrome-c were investigated by ^<13>C- and ^<19>F-NMR spectroscopy. By the complex forma- tion between OYE and NADP^4, ^<19>F-NMR signal of 8F-FMN-OYE shifted tohigher field, and remarkable changes of 4a- and 10a-^<13>C NMR signalswere observed. When cytochrome c was bound to the reconstituted enzymes, 8-^<19>F and ^<23>C NMR signals shifted to higher fields, whereas 4-, 4a-, 10a-^<13>C NMR signals shifted to lower fields. It is tempting to speculate, on the basis of these findings, that the electron transfer from aYE to cytochrome c occurs through the xylene edge of FMNsubsequent to the electron density elevation at the xylene portion upon complex formation with cytochrome c. The expression system of acyl-CoA dehydrogenase has been established to yield an abundant enzyme. By use of this system, we will get more detailed information on the reaction mechanism of the enzyme.
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