Development of the innovative technique for addition of sugar chain using microbial enzymes and its application on the syntheses of functional glycoconjugates
| Project/Area Number |
25292063
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Partial 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, 教授 (70109049)
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| Co-Investigator(Kenkyū-buntansha) |
KATAYAMA Takane 石川県立大学, 生物資源環境学部, 准教授 (70346104)
KATOH Toshihiko 石川県立大学, 生物資源環境学部, 助教 (40724612)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2015: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2014: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2013: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
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| Keywords | エンドグリコシダーゼ / 糖転移反応 / 糖鎖付加 / 糖鎖リモデリング / オキサゾリン化合物 / 免疫グロブリンＧ / コアフコース / 細胞傷害活性 / 抗体医薬 / 免疫グロブリンG / Endo-M / 部位特異的変異酵素 / イムノグロブリンG / グライコシンターゼ |
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| Outline of Final Research Achievements |
Endo-M, an endo-β-N-acetylglucosaminidase from the fungus Mucor hiemalis, is an endoglycosidase that possesses not only hydrolysis activity which liberates N-glycan leaving the innermost N-acetylglucosamine residue but also transglycosylation activity which transfers N-glycan from a donor onto a hydroxyl group-containing compounds. We generated several Endo-M mutant enzymes with transglycosidase-like activity to synthesize functional glycoconjugates. We found that N175H mutant enzyme showed glycosynthase-like activity and could synthesize a sialo-glycoform of glycoconjugates. Using this mutant, we could change the glycan of the recombinant human IgG from yeast to the sialo-glycan which is compatible to human. Moreover, Endo-M mutant enzyme W251N showed to have altered substrate specificities, exhibiting the hydrolysis activity for a core-α1,6-fucosylated glycan. This mutant could act on IgG-derived core-fucosylated glycopeptides of rituximab and that of the human lactoferrin.
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Report
(4 results)
Research Products
(23 results)
