Development of orthgonal liposomal fusion method using artificial phospholipids and its application

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K05160
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 27040:Biofunction and bioprocess engineering-related
• Research Institution: Nagoya University
• Principal Investigator: IWASAKI YUGO 名古屋大学, 生命農学研究科, 准教授 (50273214)
• Co-Investigator(Kenkyū-buntansha): 中野 秀雄 名古屋大学, 生命農学研究科, 教授 (00237348)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: リン脂質

Outline of Final Research Achievements

The following results were obtained with the aim of establishing a heterologous liposome fusion method.
(1) Artificial phospholipids with azido and alkyne groups at the polar heads were synthesized, and the click reaction was confirmed to proceed. The click reaction was performed using liposomes containing this lipids, but liposome fusion could not be confirmed. (2) For protein conjugation to the liposome surface, we synthesized an artificial phospholipid with a thioester group attached to the polar part and successfully covalently bound it to the liposome surface by reacting it with N-terminal Cys-type GFP. (3) We obtained a promising candidate that covalently binds phospholipids to the enzyme molecule itself in an autocatalytic manner, by engineering a phospholipase D, a phospholipid converting enzyme.

Free Research Field

酵素工学

Academic Significance and Societal Importance of the Research Achievements

本課題の最終目的である直交型リポソーム融合は達成できなかったが、種々の人工リン脂質の簡便合成法やチオエステル型リン脂質によるリポソーム表面へのタンパク質結合法を確立したことは学術的に意義がある。さらに、タンパク工学改変により自己触媒的にリン脂質に結合する改変型PLDの候補を取得できた。この改変酵素をさらに改良すれば、リポソームのみならず生細胞の表面に任意のタンパクを結合させることも可能になり、細胞工学において有用なツールとなる。