| Project/Area Number |
16360413
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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 |
Biofunction/Bioprocess
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| Research Institution | Kobe University |
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Principal Investigator |
KONDO Akihiko Kobe University, Faculty of Engineering, Professor, 工学部, 教授 (40205547)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUDA Hideki Kobe University, Graduate School of Science and Technology, Professor, 大学院・自然科学研究科, 教授 (30263396)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 2005: ¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 2004: ¥6,700,000 (Direct Cost: ¥6,700,000)
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| Keywords | Yeast / Cell Surface Engineering / Dockerin / Cohesin / Cellulosome / Faburication / Biomass / Cellulase |
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| Research Abstract |
To reduce the cost of immobilized enzymes, heterogonous enzymes displaying recombinant microorganisms have been developed by many researchers. However, in a current cell-surface display system to perform the complicated reaction, it is necessary to control the ration of the proteins displayed on the cell surface. To overcome this problem, we constructed a novel cell-surface display system using two pairs of proteins that assemble each other, 1 pair namely ZZ domain of protein A derived from Staphylococcus aureus and Fc part of human immunoglobulin G. And 2 pair cohesin and dockerin of cellulosome from Clostridium cellulovorans. In this system, scaffolding protein (ZZ-Coh-Coh) is displayed on the cell surface and the target protein fused to the Fc or Dockerin are secreted. Using this system, recombinant Trichoderma reesei endoglucanase II was displayed on the yeast cell surface. Display of the EGII on the cell surface was confirmed by hydrolysis experiment using β-glucan as a substrate and EGII activity was detected in the cell-pellet fraction. Following experiment showed that, EGII and Aspergillus aculeatus β-glucosidase 1 were co-displayed on the cell surface. The resulting yeast cells hydrolyzed β-glucan to glucose. This study demonstrated simultaneous display of two enzymes on the yeast cell surface can be accomplished using this cell surface display system.
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