| Project/Area Number |
10671994
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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 |
Chemical pharmacy
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| Research Institution | Health Sciences University of Hokkaido |
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Principal Investigator |
SEKIZAKI Haruo Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Associate Professor, 薬学部, 助教授 (50094834)
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| Co-Investigator(Kenkyū-buntansha) |
TANIZAWA Kazutaka Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Professor, 薬学部, 教授 (90001049)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | Chum Salmon Trypsin / Cold-Adapted Trypsin / Anionic Trypsin / Inverse Substrate / X-Ray Analysis / Enzymatic Oligopeptide Synthesis / 逆性基質 |
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| Research Abstract |
Peptide synthesis by protease-catalyzed reverse reaction has been extensively studied with a variety of model oligopeptides. It is known that enzymatic peptide synthesis is more advantageous than chemical synthesis in many respects. We demonstrated successful application of inverse substrates such as p-amidinophenyl, p-guanidinophenyl, and p-(guanidinomethyl)phenyl esteres to trypsin-catalyzed coupling.
Many studies on the characterization of trypsins from cold-adapted species have been reported These trypsins display substantially higher catalytic efficency than their manmalian counterparts. Thus, trypsin from the pyloric caeca of chum salmon (Onchorhynchus keta) is expected to be a highly efficent catalyst for enzymatic peptide synthesis.
The purpose of the this study is to elucidate the requirements for chum salmon trypsin-catalyzed peptide coupling reaction of inverse substrates with a view to utilizing the reaction as a preparative method for oligopeptide synthesis.
In 1998, we achieved the isolation and elucidation of kinetic properties of an anionic trypsin from chum salmon, and its application to enzymatic peptide synthesis using Boc-amino acid p-amidinophenyl ester as acyl donors.
In 1999, we achieved the strucure determination and refinement of benzamidine-inhibited anionic trypsin from chum salmon at 1.8 Å resolution, and application of various inverse substrates to chum salmon trypsin-catalyzed peptide synthesis. The method was useful for the preparation of various oligopeptides containing D-amlno acids and nonproteinogenic amino acids, respectively. We also achieved the preparation of N-protected Leu-enkephalin amide by this method. We prepare some reports for these results. Four articles already published part of a study on the related enzymatic peptide synthesis.
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