Clarification of cellulase and xylanase induction mechanism to direct toward enzymatic saccharification of cellulosics
| Project/Area Number |
13650851
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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 |
生物・生体工学
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| Research Institution | Nagaoka University of Technology |
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Principal Investigator |
MORIKAWA Yasushi Bioengineering, Professor, 工学部・生物系, 教授 (50239638)
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| Co-Investigator(Kenkyū-buntansha) |
OGASAWARA Wataru Bioengineering, Assistant Professor, 工学部・生物系, 助手 (40292172)
OKADA Hirofumi Bioengineering, Associate Professor, 工学部・生物系, 助教授 (70233343)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Cellulase / Xylanase / Induction / Promoter / Trichoderma reesei / Homologous recombination / Gene / Trichoderma reesei / 相同組み換え / 誘導 / xyn3 / eg13 / 転写活性化因子 |
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| Research Abstract |
Xylanase III in Trichoderma reesei has been found to be co-induced with the cellulases and not to be induced by xylan and its derivatives which are general inducers in xylanase expression. Moreover, the enzyme gene (xyn3) was expressed in T. reesei PC-3-7 but not in T. reesei QM9414, the parent strain of the former, although both strains have the same chromosomal gene and are able to express the cellulase genes similarly.
Firstly, the nucleotide sequences of the promoter region of the genes of both strains were determined to clear the regulatory mechanism of cellulase and xylanase induction in T. reesei. The both sequences were completely the same, and the putative binding region with the known regulatory factors, cellulase gene-relating ACE I, ACE II and catabolite repressor Cre I, were presented in the promoter, suggesting that one or some unknown regulatory factors were deleted in T. reesei QM9414 or PC-3-7.
Next, the promoter region (ca. 3. 4 kbp) of endoglucanase III gene (eg13) was cloned and sequenced, which is expressed extremely lower than are the other main cellulase genes such as cellobiohydrolase I and II genes in T. reesei. Furthermore, the homologous transformation system in T. reesei was confirmed using amdS gene as a maker.
From these results, the deleted promoter regions of xyn3 and eg13 followed by the reporter gene (α-glucronidase gene) were constructed, and the in vivo reporter assay are now being tried in T. reesei transformed with the deleted promoter regions using the homologous recombination system. On the other hand, in vivo interactions between the promoters and nuclear proteins containing reguratory factors are also being investigated using band-shift assays.
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Report
(3 results)
Research Products
(6 results)
