Molecular Analysis of Carbohydrases Showing Different Reaction by Novel Structure and Its Application
| Project/Area Number |
17380060
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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 |
Applied biochemistry
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| Research Institution | Hokkaido University |
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Principal Investigator |
KIMURA Atsuo Hokkaido University, Research Faculty of Agriculture, Professor (90186312)
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| Co-Investigator(Kenkyū-buntansha) |
MORI Haruhide Hokkaido University, Research Faculty of Agriculture, Associate Professor (80241363)
OKUYAMA Masayuki Hokkaido University, Research Faculty of Agriculture, Assistant Professor (00344490)
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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 |
¥15,890,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2007: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2006: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2005: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Enzyme reaction / Sugar / Protein engineering |
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| Research Abstract |
This project is about the enzymes having the structure similar to α-glucosidase; i.e. α-xylosidase, glucan lyase, cyclic-tetrasaccharide-forming enzyme, and α-glucosidase, each of which catalyzes the different reaction. Three-dimensional structure available recently allows us to analyze the molecular mechanism of reactions exhibited by four enzymes. The purposes of research are 1) to elucidate the relationship between substrate and amino acid residue(s) in the catalytic site; 2) to analyze the function of catalytic residues; 3) to elucidate the structural element(s) to display the above-described different reactions; 4) to synthesize the useful enzyme. Results are as follows.
(1) We analyzed the amino acid residues in the catalytic site of α-xylosidase to recognize α-xyloside-structure, and succeeded in conversion of α-xylosidase into α-glucosidase by the mutagenesis of its structural elements. (2) The catalytic residues were identified and their functions were investigated. (3) Amino acid replacement of α-glucosidase (a hydrolyzing enzyme) lost its hydrolytic activity and enhanced the transglucosidation ability, meaning the conversion of hydrolyzing enzyme into transferring enzyme. (4) We have succeed in change of transferring products.
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Report
(4 results)
Research Products
(118 results)
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[Journal Article] Rice α-glucosidase isozymes and isoforms showing different starch granules-binding and -degrading ability2008
Author(s)
Nakai H, Tanizawa S, Ito T, Kamiya K, Yamamoto T, Matsubara K, Kim YM, Sakai M, Sato H, Imbe T, Okuyama M, Mori H, Sano Y, Chiba S, Kimura A
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Journal Title
Biocatal Biotransfor 104-110
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Multiple forms of α-glucosidase in rice seeds (Oryza sativa L, var Nipponbare)2007
Author(s)
Nakai H, Ito T, Hayashi M, Kamiya K, Yamamoto T, Matsubara K, Kim YM, Wongchawalit J, Okuyama M, Mori H, Chiba S, Sano Y, Kimura A
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Journal Title
NAID
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] A novel cyclic isomaltooligosaccharide (cycloisomaltodecaose, CI-10) produced by Bacillus circulans T-3040 displays remarkable inclusion ability compared with cyclodextrins2007
Author(s)
Funane K, Terasawa K, Mizuno Y, Ono H, Miyagi T, Gibu S, Tokashiki T, Kawabata Y, Kim YM, Kimura A, Kobayashi M
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Molecular cloning of a cDNAs for three α-glucosidases from European honeybee, Apis mellifera L2007
Author(s)
Nishimoto M, Mori H, Moteki T, Takamura Y, Iwai G, Wongchawalit J, Surarit R, Svasti J, Kimura A, Chiba S
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Journal Title
Biosci Biotechnol Biochem 1703-1716
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Molecular cloning of cDNA for trehalase from European honeybees, Apis mellifera L, and its heterologous expression in Pichia pastoris2007
Author(s)
Lee JH, Saito S, Mori H, Nishimoto M, Okuyama M, Kim D, Wongchawalit J, Kimura A, Chiba S
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Journal Title
Biosci Biotechnol Biochem 2256-2265
NAID
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Function-unknown glycoside hydrolase family 31 proteins expressed in rice ripening and germinating stages are α-glucosidase and α-xylosidase2007
Author(s)
Nakai H, Tanizawa S, Ito T, Kamiya K, Yamamoto T, Matsubara K, Kim YM, Sakai M, Sato H, Imbe T, Okuyama M, Mori H, Sano Y, Chiba S, Kimura A
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Journal Title
Description
「研究成果報告書概要(欧文)」より
Related Report
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[Journal Article] Enzymatic synthesis and characterization of arbutin glucosides using glucansucrase from Leuconostoc mesenteroides B-1299CB2007
Author(s)
Moon YH, Nam SH, Kang J, Kim YM, Lee JH, Kang HK, Breton V, Jun WJ, Park KD, Kimura A, Kim D
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Journal Title
Appl Microbiol Biot 559-567
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Purification and characterization of α-glucosidase I from Japanese honeybee (Apiscerana japonica), and molecular cloning of its cDNA2006
Author(s)
Wongchawalit J, Yamamoto T, Nakai H, Kim Y-M, Sato N, Nishimoto M, Okuyama M, Mori H, Saji O, Chanchao C, Wongsiri S, Surarit R, Svasti J, Chiba S, Kimura A
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Journal Title
Biosci Biotechnol Biochem 2889-2898
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Interactions between barley α-amylases, substrates, inhibitors and regulatory proteins2006
Author(s)
Hachem MA, Bozonnet S, Willemoes M, Bonsager BC, Nielsen MM, Fukuda K, Kramhoft B, Maeda K, Sigurskjold BW, Hahlund P, Finnie P, Mori H, Robert X, Jensen MH, Tranier S, Aghajari N, Haser R, Scensson B
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NAID
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Binding of carbohydrates and protein inhibitors to the surface of α-amylases2005
Author(s)
Bozonnet S, Bonsager BC, Kramhoft B, Mori H, Hachem MA, Willemoes M, Jensen MT, Fukuda K, Nielsen PK, Juge N, Aghajari N, Tranier S, Robert X, Haser R, Scensson B
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Description
「研究成果報告書概要(欧文)」より
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