Biochemical preparation of isotpe-labelled lipopolysaccharide and nmr analysis of host recognition mechanism of phage

2016 Fiscal Year Final Research Report

• Project/Area Number: 26660085
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: Mie University
• Principal Investigator: Inagaki Minoru 三重大学, 生物資源学研究科, 教授 (20242935)
• Research Collaborator: KARITA Shuichi 三重大学, 大学院生物資源学研究科, 教授 (90233999) ; TAKEDA Shigeki 群馬大学, 大学院理工学府, 教授 (80282854) ; FUJITA Katsutoshi 独立行政法人・製品評価技術基盤機構NITE, 資源保存課主任
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: リポ多糖 / 糖転移酵素 / 相同組み換え / 変異株 / 糖鎖構造解析 / 電気泳動 / 質量分析

Outline of Final Research Achievements

For incorpolation of 13C isotope-labelled galactose into the non-reducing end of lipopolysaccharide core structure, galE gene on E. coli C bacterium was determined on the gene. The mutant of E. coli F2513 were prepared by deletion of Galactosyl transferase genes, waaW and waaT. However, those mutants doesn't propagete in the cultural medium, and not yet certified the altered saccharide stracture on R-core lipopolysaccharide. By using the ASKA(-) clone, JW3598AM, the protein fraction of WaaU (hetosyl transferase) was prepared and submitted to treat the Salmonella bacterium. The incorpolation of an unnatural Hep residue was confirmed on the lipopolysacchride of Salmonella, using DOC-PAGE and LC-ESI-MS.

Free Research Field

生物有機化学