Innovative technique for production of biomedicines with uniform sugar chains by use of microbial enzymes
| Project/Area Number |
16K07698
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied biochemistry
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| Research Institution | Ishikawa Prefectural University |
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Principal Investigator |
Yamamoto Kenji 石川県立大学, 生物資源環境学部, 特任教授 (70109049)
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| Co-Investigator(Kenkyū-buntansha) |
加藤 紀彦 京都大学, 生命科学研究科, 助教 (40724612)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | バイオ医薬品 / 糖鎖 / エンドグリコシダーゼ / 糖転移反応 / リモデリング / インターフェロン-γ / インターロイキン-３ / Pichia pastoris / エンドーＭ / インターロイキン３ / インターフェロンーγ / エンド-M / インターロイキン-3 / エンド-Ｍ |
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| Outline of Final Research Achievements |
As the results of this work, we succeeded in the gene cloning and expression of glycoproteins such as human Interferon-gamma and Interloikin-3 which are known as the biomedicine, by use of Pichia yeast as a host. The recombinant glycoproteins obtained were treated with Endo-H to release their high-mannose type sugar chains. Thereafter the addition of human compatible sialo-complex type sugar chains of glycopeptides derived from hen egg yolk to these sugar chain-depleted proteins was carried out using the transglycosylation activity of the mutant enzyme of mold endoglycosidase (Endo-M) and the exchange (remodeling) of sugar chain was performed. It was intended to create human compatible biomedicines. However we could obtain only a little amount of the purposed products. Then in order to effectively act to the substrate, the minimization of the size of Endo-M was attempted, but it did not succeed.
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| Academic Significance and Societal Importance of the Research Achievements |
バイオ医薬品の多くは動物細胞を用いて調製されているが、多量生産が困難であり、糖タンパク質であるためにタンパク質は同一であっても薬効に影響を与える糖鎖の構造が不均一である故に同じ医薬品でも糖鎖構造の違いによって薬効に差が生じる。本研究の成果によって、多量生産できる微生物の酵母を用いてヒト由来サイトカインなどのバイオ医薬品の多量調製が可能であることが明らかになった事実は今後のバイオ医薬品の製造に有用な意義を持つ。さらに本研究では充分な成果を得ることができなかったが、本研究が微生物の酵素を用いたリモデリング法によって均一な糖鎖をもつバイオ医薬品を多量調製する技術開発に示唆を与えた学術的意義は大きい。
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Report
(4 results)
Research Products
(22 results)
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[Journal Article] Chemo-enzymatic synthesis of the glucagon containing N-linked oligosaccharide and its characterization.2018
Author(s)
Higashiyama, T., Umekawa, M., Nagao, M., Katoh, T., Ashida, H., Yamamoto, K.
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Journal Title
Carbohydrate Research
Volume: 455
Pages: 92-96
DOI
Related Report
Peer Reviewed
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[Journal Article] Glycosylation engineering of therapeutic IgG antibodies: challenges for the safety, functionality and efficacy2018
Author(s)
Mimura, Y., Katoh, T., Saldova R, O’Flaherty, R., Izumi, T., Mimura-Kimura, Y., Utsunomiya, T., Mizukami, Y., Yamamoto, K., Matsumoto, T., Rudd, PM.
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Journal Title
Protein Cell.
Volume: 9（1）
Issue: 1
Pages: 47-62
DOI
NAID
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Specificity of donor structures for endo-beta-N-acetylglucosaminidase-catalyzed transglycosylation reactions.2018
Author(s)
Ishii, N., Ogiwara, K., Sano, K., Kumada, J., Yamamoto, K., Matsuzaki, Y., Matsuo, I.
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Journal Title
ChemBioChem,
Volume: 19
Issue: 2
Pages: 136-141
DOI
Related Report
Peer Reviewed
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[Journal Article] Generation of a mutant Mucor hiemalis endoglycosidase that prefers core-fucosylated N-glycans.2016
Author(s)
Katoh, T., Katayama, T., Tomabechi, Y., Nishikawa, Y., Kumada, J., Matsuzaki, Y., Yamamoto, K
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Journal Title
Journal of Biological Chemistry
Volume: 291
Issue: 44
Pages: 23305-23317
DOI
Related Report
Peer Reviewed / Open Access / Acknowledgement Compliant
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[Presentation] 微生物の応用から見た糖鎖2016
Author(s)
山本憲二
Organizer
糖鎖科学中部拠点、第13回「若手の力」フォーラム
Place of Presentation
岐阜大学（岐阜県・岐阜市）
Year and Date
2016-11-26
Related Report
Invited
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