| Project/Area Number |
06304015
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| Research Category |
Grant-in-Aid for Co-operative Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
TONOMURA Ben'ichiro Kyoto Univ., Fac.of Agric., Prof., 農学部, 教授 (20026545)
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| Co-Investigator(Kenkyū-buntansha) |
SAKIYAMA Fumio Osaka Univ., Protein Res.Inst., Prof., 蛋白質研究所, 教授 (40029947)
ODA Jun-ichi Kyoto Univ., Inst.Chem.Res., Prof., 化学研究所, 教授 (50027041)
IWANAGA Sadaaki Kyushu Univ., Fac.of Sci., Prof., 理学部, 教授 (90029942)
IMOTO Yaiji Kyushu Univ., Fac.of Pharm., Prof., 薬学部, 教授 (90038282)
ASHIDA Tamaichi Nagoya Univ., Fac.of Enging., Prof., 工学部, 教授 (10029936)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥30,400,000 (Direct Cost: ¥30,400,000)
Fiscal Year 1995: ¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 1994: ¥15,400,000 (Direct Cost: ¥15,400,000)
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| Keywords | Protein Engineering / Structure of Protein / X-ray Crystallography of Protein / Heat Stability of Protein / Base Sequence of Gene / Substrate Specificity of Enzyme / NMR of Protein / Chemical Modification of Protein / 遺伝子の塩基配列 / タンパク質の耐熱性 / タンパク質間特異的分子認識 / 部位特異的変異 |
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| Research Abstract |
The structures of plant trypsin inhibitors, their complexes with trypsin, snake venom lipase A2, glutathione synthetase and aspartic acid synthetase of E.coli resolved by X-ray crystallography, and their reaction mechanisms analyzed.
Mechanisms of heat denaturation and polypeptide folding of lysozyme, and alteration of the reaction mechanism elucidated by protein engineering methods. The binding of blood coagulation VIIa factor, and activation mechanism of a serine protease in the cagulation of horse-shoe crab serum investigated. Chemical modification of glutathione synthetase with PEG investigated for clinical application. Solution structure of the single-chained monelline elucidated by NMR.The high substrate specificity of Acromobacter protease I analyzed with recombinant enzymes. Accumulative heat resistance of glucosidases by multiple introduction of Pro residues proved. The nucleotide sequence of lysyl tRNA synthetase gene of B.stearothermophilus elucidated and the binding of substrates kinetically analyzed. A hybrid enzyme between two dioxygenases showed broader substrate specificity. Chemical modification of lysozyme with novel dextran and PEG derivatives resulted in the increase of stability and reactivity of the enzyme. Six bacterial peptidases specific for Pro residue purified, the genes of the enzymes cloned, the nucleotide sequences elucidated, over-expressed, and the enzymes were crystallized.
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