Precise hybrid synthesis of glycoprotein through amino acid sequence-specific introduction of oligosaccharide followed by enzymatic transglycosylation reaction
Project/Area Number |
22550105
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Polymer chemistry
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Research Institution | Tohoku University |
Principal Investigator |
KOBAYASHI Atsushi 東北大学, 工学(系)研究科(研究院), 助教 (90361138)
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Project Period (FY) |
2010-04-01 – 2014-03-31
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Project Status |
Completed (Fiscal Year 2013)
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Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2013: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2012: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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Keywords | 糖タンパク質合成 / 酵素化学合成 / アマドリ転移反応 / 配列特異的反応 / 高分子反応 / アマドリ転位反応 / 酵素的配糖化 / 糖オキサゾリン / 活性化糖 / 複合糖質 / 糖タンパク質 / 化学-酵素法 / リゾチーム / 基質エンジニアリング / 加水分解酵素 / 酵素的糖転移反応 / 受容体特異性 / N-結合型糖鎖 / 無保護糖活性化 / 糖蛋白質 / 一段階糖誘導体合成 / 糖蛋白質糖鎖 |
Research Abstract |
It is difficult to obtain homogeneous glycoproteins, although it is increasing our knowledge concerning physiological functions of glycoproteins. Nowadays, it is very easy to obtain bare proteins that fold appropriately by using prokaryote cells like Escherichia coli, and it is possible to obtain desired glycoproteins by conjugating with oligosaccharide and proteins precisely. Recently, it was reported that the Amadori rearrangement occurred between oligosaccharides and proteins in a sequence specific manner. Based on this report, I have investigated the effect of additives on the Amadori rearrangement reaction and acceleration of its reaction rate. The most striking result was that the effects of borate ion were depended upon chemical structure of sugars. For example, very large reaction rate using D-fructose was observed in phosphate buffer, although almost no reaction was observed in borate buffer. Investigation of sequence specificity of the Amadori rearrangement is under way.
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Report
(5 results)
Research Products
(55 results)
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[Journal Article] Crystal Structures of Glycoside Hydrolase Family 51 α-L-Arabinofuranosidase from Thermotoga maritima2012
Author(s)
Do-Hyun Im, Kei-ichi Kimura, Fumitaka Hayasaka, Tomonari Tanaka, Masato Noguchi, Atsushi Kobayashi, Shin-ichiro Shoda, Kentaro Miyazaki, Takayoshi Wakagi, Shinya Fushinobu
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Journal Title
Bioscience, Biotechnology, and Biochemistry
Volume: 76
Issue: 2
Pages: 423-428
DOI
NAID
ISSN
0916-8451, 1347-6947
Related Report
Peer Reviewed
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[Journal Article] 4,6-Dimethoxy-1,3,5-triazin-2-ylβ-D-glycosaminides : Novel Substrates for Transglycosylation Reaction Catalyzed by Exo-β-D-glucosaminidase from Amycolatopsis orientalis2011
Author(s)
Tomonari Tanaka, Tomonori Wada, Masato Noguchi, Masaki Ishihara, Atsushi Kobayashi, Takayuki Ohnuma, Tamo Fukamizo, Ryszard Brzezinski, Shin-ichiro Shoda
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Journal Title
Journal of Carbohydrate Chemistry
Volume: 31
Issue: 8
Pages: 634-646
DOI
Related Report
Peer Reviewed
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[Journal Article] Efficient transfer of sialo-oligosaccharide onto proteins by combined use of a glycosynthase-like mutant of Mucor hiemalis endoglycosidase and synthetic sialo-complex-type sugar oxazoline2010
Author(s)
Midori Umekawa, T. Higashiyama, Y. Koga, T. Tanaka, M. Noguchi, Atsushi Kobayashi, S. Shoda, Wei Huang, Li-Xi Wang, Hisashi. Ashida, and Kenji Yamamoto
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Journal Title
Biochimica et Biophysica Acta, General Subjects
Volume: 1800
Issue: 11
Pages: 1203-1209
DOI
NAID
Related Report
Peer Reviewed
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[Journal Article] 4,6-Dimethoxy-1,3,5-triazine oligoxyloglucans : Novel one-step preparable substrates for studying action of endo-β-1,4-glucanase III from Trichoderma reesei2010
Author(s)
Atsushi Kobayashi, Tomonari Tanaka, K. Watanabe, Masaki Ishihara, Masato Noguchi, H. Okada, Yasushi Morikawa, and Shin-ichiro Shoda
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Journal Title
Bioorganic and Medicinal Chemistry Letters
Volume: 20
Issue: 12
Pages: 3588-3591
DOI
Related Report
Peer Reviewed
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[Journal Article] 4,6-Dimethoxy-1,3,5-triazine oligoxyloglucans : Novel one-step preparabie substrates for studying action of endo-β-1,4-glucanase III from Trichoderma reesei2010
Author(s)
A.Kobayashi, T.Tanaka, K.Watanabe, M.Ishihara, M.Noguchi, H.Okada, Y.Morikawa, S.-i.Shoda
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Journal Title
Bioorg.Med.Chem.Lett.
Volume: 20
Pages: 3588-3591
Related Report
Peer Reviewed
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[Journal Article] Efficient transfer of sialo-oligosaccharide onto proteins by combined use of a glycosynthase-like mutant of Mucor hiemalis endoglycosidase and synthetic sialo-complex-type sugar oxazoline2010
Author(s)
M.Umekawa, T.Higashiyama, Y.Koga, T.Tanaka, M.Noguchi, A.Kobayashi, S.-i.Shoda, W.Huang, L-X.Wang, H.Ashida, K.Yamamoto
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Journal Title
Biochim.Biophys.Acta, Gen.Subj.
Volume: 1800
Pages: 1203-1209
NAID
Related Report
Peer Reviewed
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[Presentation] A Novel Chemo-Enzymatic Synthesis αGlcNAC-linked Oligosaccharides2010
Author(s)
Akiko Tsuchida, Masaya Fujita, Miwa Nakamura, Tomonari Tanaka, Kotaro Goto, Kenji Osumi, Yoko Hirose, Hisashi Ashida, Kenji Yamamoto, Masato Noguchi, Atsushi Kobayashi, Shin-ichiro Shoda, Mamoru Mizuno
Organizer
International Carbohydrate Symposium
Place of Presentation
幕張メッセ(千葉)
Year and Date
2010-08-02
Related Report
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