| Project/Area Number |
09044214
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | NAGOYA UNIVERSITY |
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Principal Investigator |
TSUKAGOSHI Norihiro NAGOYA UNIVERSITY,DEPARTMENT OF AGRICULTURE,PROFESSOR, 農学部, 教授 (50115599)
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| Co-Investigator(Kenkyū-buntansha) |
HEMMING F.W. ノッチンガム大学, 生化学部, 教授
PEBERDY J.F. ノッチンガム大学, 生命科学部, 教授
KATO Masashi NAGOYA UNIV., DEPT.OF AGRICULTURE,ASSISTANT PROFESSOR, 農学部, 助手 (70242849)
KOBAYASHI Tetsuo NAGOYA UNIV., DEPT.OF AGRICULTURE,ASSOCIATE PROFESSOR, 農学部, 助教授 (20170334)
JOHN F. Peberdy NOTTINGHAM UNIV., DEPT.OF LIFE SCIENCE,PROFESSOR
FRANK W. Hemming NOTTINGHAM UNIV., DEPT.OF BIOCHEMISTRY,PROFESSOR
PEBARDY J.F. ノッチンガム大学, 生命科学部, 教授
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1997: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | A.oryzae / A.nidulans / Glycan structure of Taka-amylase A, (TAA) / Glycan structure of AoTAA / Glycan structure of AnTAA / Difference in glycosylation between A.oryzae and A.nidulans / タカミラーゼA(TAA)糖鎖 |
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| Research Abstract |
In order to elucidate the significance of protein glycosylation during protein secretion and in expression of enzymatic function, the glycosylation mechanism in filamentous fungi has been investigated molecular-biologically and biochemically using Taka-amylase A as a model enzyme. Our joint works carried out at both sites during last two years are summarized as follows ;
1) The A.oiyzae Taka-amylase A (AoTAA) and Taka-amylase A produced by A.nidulans carrying the A.ozyzae Taka-amylase A gene (AnTAA) were highly purified to homogeneity from commercially available enzyme preparation and the culture broth of an A.nidulans trasformant, respectively.
2) The glycan structures of AoTAA and AnTAA were analyzed by lectin blotting. Both enzymes contained mannose as a component of gylcans. However, a highly glycosylated protein with a low molecular mass was found to be contaminated in those preparations during this analysis. This low-molecular-mass glycoprotein interfered further monosaccharide analysis of the glycans. Although we made our best to get rid of this contaminant from AnTAA preparations using several purification methods, we have not yet succeed to remove the contaminant from the enzymes.
3) The glycan structures were analyzed by fluorescent labeling technique and we found that AoTAA and AnTAA comprised 7 and S monosaccharides, respectively. This indicates the glycosylation process is different in A.oryzae and A.nidulans
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