| Project/Area Number |
13480187
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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 |
Bioorganic chemistry
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
HIRATAKE Jun Kyoto Univ., Inst.for Chem.Res., Assoc.Prof., 化学研究所, 助教授 (80199075)
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| Co-Investigator(Kenkyū-buntansha) |
TAKADA Masayasu Nihon Shokuhin Kako Co., Ltd., Investigator, 研究所, 研究員
MIZUTANI Masaharu Kyoto Univ., Inst.for Chem.Res., Instructor, 化学研究所, 助手 (60303898)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2001: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Keywords | β-Glycosylamidines / β-Glycosidase inhibitors / Substrate analogs / Glycon substrate specificity / Aglycon substrate specificity / Affinity adsorbent / Tea leaf β-glucosidases / β-Galactosidases / グリコシダーゼ / グリコシルアミジン誘導 / 基質特異的グリコシダーゼ阻害剤 / グリコン特異性 / アグリコン特異性 / グリコシダーゼ精製 / グリコシルアミジン誘導体 / ワンステップ精製 |
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| Research Abstract |
This study aims to develop novel glycosidase inhibitors and to use these inhibitors as versatile research tools in glycosidases studies from a bioorganic point of view. For this purpose, we have designed and synthesized f -glycosylamidine derivatives as highly potent and glycon-selective inhibitors of β-glycosidases. β-Glycosylamidines are a substrate analog in which a positively charged amidino group is incorporated into a sugar surrogate in such a way as to maintain the whole structure of the glycon, including the stereochemistry at C-1. The β-glycosylamidines are readily synthesized from various sugars n two steps without using any protective groups. The β-glycosylamidines with each glycon moiety serve as highly potent inhibitors (K_i<0.1μM) toward the β-glycosidases with the corresponding glycon substrate specificity, while the β-glycosidases with different glycon substrate specificity are not inhibited significantly. Furthermore, varying structures can be introduced into the "agly
… More
con" moiety of the inhibitor to increase or tune the inhibition potency and selectivity towards various β-glycosidases. Therefore the β-glycosyamidines are versatile β-glycosidase inhibitors in which both the glycon and aglyon moieties can be adjusted according to the enzyme to be inhibited. This property can be successfully used as a ligand for affinity chromatography of glycosidases.. Thus, tea leaf β-glucosidases were purified to homogeneity in one step by the affinity adsorbent with β-glucosylamidine as ligand, while a β-galactosidase from mold was also purified in one step by an affinity chromatography with β-galactosylarnidine as ligand. Other β-glycosidases such as β-primeverosidase, a novel diglycoside-specific glycosidase, was also affinity-purified successfully by using the corresponding glycosylamidine as ligand. The purified tea leaf β-glucosidases were subjected to amino acid sequence analysis to clone the genes. Hence the β-glycosylamidines are versatile research tools in glycosidase studies spanning from the isolation, the characterization and the gene cloning. Less
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