Development of the innovative technique for addition of sugar chain using microbial enzymes and its application on the syntheses of functional glycoconjugates

2015 Fiscal Year Final Research Report

• Project/Area Number: 25292063
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Ishikawa Prefectural University
• Principal Investigator: YAMAMOTO Kenji 石川県立大学, 生物資源環境学部, 教授 (70109049)
• 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)
• Project Period (FY): 2013-04-01 – 2016-03-31
• 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

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.

Free Research Field

応用微生物学