Manipulation of bacterial membrane vesicle for infection control applications

Research Project

Project/Area Number	25670211
Research Category	Grant-in-Aid for Challenging Exploratory Research
Allocation Type	Multi-year Fund
Research Field	Bacteriology (including mycology)
Research Institution	Osaka University
Principal Investigator	TOBE Toru 大阪大学, 医学(系)研究科(研究院), 教授 (70207596)
Project Period (FY)	2013-04-01 – 2016-03-31
Project Status	Completed (Fiscal Year 2015)
Budget Amount *help	¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000) Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000) Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000) Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywords	外膜小胞 / デリバリーシステム / エンドトキシン / LPS / OMV / ターゲッティング / 物質輸送 / 薬剤耐性因子 / 腸内環境因子 / 抗生物質
Outline of Final Research Achievements	Almost all bacteria are known to produce membrane vesicle. Outer membrane vesicle is a membrane vesicle produced from outer membrane of gram-negative bacteria. In this project, we aimed to strengthen basic knowledge on OMV characteristics for future application. We used E. coli as a model and obtained the following results. We found that production efficiency of OMV was affected by variety of environmental factors and different among strains, and that only small amount of cytoplasmic proteins were included in OMV. By using kanamycin-resistance as a marker protein, transfer of kanamycin-inactivation enzyme from resistant strain to sensitive strain was suggested. On the other hand, we developed the system to present any protein fragment on cell surface by using an autotransporter protein. Finally, we found the gene that modifies Lipid A and reduces endotoxin activity. These findings are useful for development of OMV-based strategy for infection control.