1998 Fiscal Year Final Research Report Summary
Development of High Efficiency Protein Refolding Processes Using Molecular Chaperons.
| Project/Area Number |
09555238
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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 | 展開研究 |
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| Research Field |
反応・分離工学
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| Research Institution | Kobe University |
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Principal Investigator |
FUKUDA Hideki Kobe University Graduate School of Science and Technology Professor, 自然科学研究科, 教授 (30263396)
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| Co-Investigator(Kenkyū-buntansha) |
OHTUKA Koutaro Nagase & Co., LTD Research and Development Center Manager, 研究開発センター, 主任研究員
KONDO Akihiko Kobe University Faculty of Engineering Associate Professor, 工学部, 助教授 (40205547)
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| Project Period (FY) |
1997 – 1998
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| Keywords | Molecular chaperon / Chaperonin / GroEL / Protein refolding / Fusion protein / Affinity separation / Immobilization |
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| Research Abstract |
Molecular chaperons are powerful to improve the refolding yield of proteins from inclusion bodies and the reactivation of thermally inactivated enzymes in various bioprocesses. In this study, to develop the high efficiency protein refolding systems based on molecular chaperons, the properties of various types of chaperonins from different organisms (i.e. Esherichia coli, Thermu sthermophilus and Bacillus stearothermophilus) were investigated. The overexpression and purification systems of these chaperonins were constructed to produce chaperonins efficiently. To enhance the reusability of chaperonins, various immobilization methods of chaperonins were tested. As the support materials, cellulose gel beads and colloidal polymer particles were adopted. Especially, chaperonins with affinity tag (His)_6 or maltose binding protein (fusion chaperonin) were found to be effective for efficient production and immobilization using affinity adsorbents. In addition, protein refolding system using chaperonin in combination with ultrafiltration system was found to be effective. Based on these results, several efficient protein refolding systems using chaperonins were proposed.
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