Synthesis of neoglycoproteins using oligosaccharide-transfer Activity with immobilized endoglycosidases

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K19090
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 38:Agricultural chemistry and related fields
• Research Institution: Kyushu University
• Principal Investigator: Takegawa Kaoru 九州大学, 農学研究院, 教授 (50197282)
• Project Period (FY): 2021-07-09 – 2024-03-31
• Keywords: エンドグリコシダーゼ / ネオグリコプロテイン / 固定化酵素

Outline of Final Research Achievements

We investigated efficient deglycosylation techniques by immobilizing α-L-fucosidase and endoglycosidase (ENGase), which are necessary for deglycosylation of sugar chains from glycoproteins. Both enzymes were found to retain their activity after immobilization on NHS-Activated Agarose. LC/MS measurements confirmed deglycosylation of IgG using both immobilized enzymes. We searched for a novel ENGase capable of deglycosylation of multi-branched glycans of human-derived glycoproteins. We expect that Barnesiella intestinihominis possesses ENGases to facilitate the utilization of complex-type N-glycans from host cells.

Free Research Field

応用微生物学

Academic Significance and Societal Importance of the Research Achievements

現在市販されているバイオ医薬品の多くが糖タンパク質であるが、最も高い生物活性を示す糖鎖構造については未だ明らかにされていない。そこで、本研究では煩雑な工程であったエンドグリコシダーゼの糖鎖転移反応による糖タンパク質均一糖鎖変換技術を固定化酵素を用いて行うことを目指した。条件検討の結果、固定化酵素による糖タンパク質の脱糖鎖は効率良く行うことが可能になった。