| Project/Area Number |
17380102
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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 |
林産科学・木質工学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SAMEJIMA Masahiro The University of Tokyo, Graduate School of Agricultural and Life Sciences, professor (30162530)
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| Co-Investigator(Kenkyū-buntansha) |
KIYOHIKO Igarashi The University of Tokyo, Graduate School of Agricultural and Life Sciences, Assistant professor (80345181)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥15,840,000 (Direct Cost: ¥15,300,000、Indirect Cost: ¥540,000)
Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2005: ¥11,700,000 (Direct Cost: ¥11,700,000)
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| Keywords | Genome / Glycoside Hydrolase / Biomass / Proteome / Cellulose / Basidiomycetes / Phanerochaete chrysosporium / Wood Rotting Fungi / Phanerochaete chtysospor / 酵素 |
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| Research Abstract |
In this project, proteomic analysis of extracellular enzymes produced by the Basidiomycete Phanerochaete chrysosporium during cellulose degradation was performed. Two-dimensional gel electrophoresis for extracellular solution, LS-MS/MS analysis for each enzyme separated on the gel to identify the partial sequence of amino acids, and searching on the P.chrysosporium database to find the corresponding genome sequences were applied for this purpose. After these processes, the cDNAs of the target enzymes depending on the purposes desired were cloned and transformed in the yeast Pichia pastoris with expression vectors to produce the recombinant enzymes of them.
On the basis of these techniques established, we have succeeded to obtain several cellulases and related enzymes produced during cellulose degradation by this fungus as recombinant enzymes. Among them, an endoglucanase in GH family 45 and a xyloglucanaase in GH family 74 were identified as novel enzymes and well characterized by this project. In addition, the novel beta-1, 3-glucanase in GH family 16, named Lam 16A, has been found during cellulose degradation by this fungus. As a collaboration work with the research group in the Swedish University of Agricultural and Life Sciences, functional and structural analysis of Lam16A was accomplished in a part of this project. Moreover, we have also succeeded to synthesis the novel cycloglucan using a site-directed mutant enzyme derived from Lam16.
In conclusion, we have established the total technical system from proteomic analysis of extracellular solution of the basidiomycete P.chrysosporium to production of recombinant enzymes for the conversion of cellulosic biomass.
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