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Microbial production of oligosialic acid

Research Project

Project/Area Number 07455327
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 生物・生体工学
Research InstitutionNagoya University, School of Engineering

Principal Investigator

IIJIMA Shinji  Department of Biotechnology, Nagoya University, School of Engineering Professor, 工学部, 教授 (00168056)

Co-Investigator(Kenkyū-buntansha) MIYAKE Matsuhide  Department of Biotechnology, Nagoya University, School of Engineering Research A, 工学部, 助手 (90252254)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1996: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1995: ¥4,600,000 (Direct Cost: ¥4,600,000)
Keywordssialic acid selectin / microbial production / sugar technology / metastasis / phage / phage / suger technology
Research Abstract

Analyzes and application of sugar in biological process have not been studied extensively. In this regard, we studied microbial production of functional sugar such as oligosialic acids and sialyl lactose, an analog of sialyl Lewis sugar.
Seven different bacteriophages were isolated from sewage with Escherichia coli K1 strains as host bacteria. These phages showed specific degrading activity for the host capsular polysaccharide, alpha-2,8-linked polysialic acid. With respect to the minimum substrate size, each enzyme had its own specificity. Of the seven isolated phages, four enzyme showed novel endo-N-acetylneuraminidase activities. We purified one of the enzyme from both phage particle and soluble fraction of infected microbial culture. We also develop an antibody against soluble neuraminidase. In addition to this, the neuraminidase gene was cloned in E.coli and the recombinant protein was purified. By immunoblotting, these 3 neuraminidase species were immunologically identical and their molecular weigh was 90kD.
By using normal endothelial cells, we were able to detect inhibitory effects of type specific polysaccharides from Streptococcus agalactiae on adhesion of cancer cells to endothelial cells, which is an essential step of cancer metastasis. The inhibition was probably due to specific structures of the bacterial polysaccharides, since the structures of the saccharides are very similar to those of cancer specific sialyl Lewis carbohydrates (sialyl Le^a and Le^x) which bind to ELAM-1 of endothelial cells.
In S.agalactiae strain specific capsular sugar, a ligand (sialyl lactose) was blanched from main chain (repeating units of Gal-GluNAc). The branchings are observed every repeating unit of main chain. We tried to digest the bacterial sugar with a beta-galactosidase and produced sialyl lactose oligomer.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] 飯島信司: "Immortalization of human endothelial cells by temperature sensitive simian virus 40" Animal Cell Technology : Developments towards the 21st century. 57-61 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 飯島信司: "Immortalization of human endothelial cells by temperature sensitive simian virus 40" The Sixth Annual Meeting of Japanese Association for Animal Cell Technology (JAACT'93) Program & Abstracts. 46- (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 飯島信司: "Isolation of immortal human endothelial cells" Bioscience, Biotechnology, and Biochemistry. 59. 912-914 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 飯島信司: "Blocking adhesion of cancer cells to endothelial cell types by S. agalactiae type-specific polysaccharides" Cytotechnology. 22. 205-210 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 飯島信司: "細菌表面多糖が示すガン細胞接着阻害活性" 化学工学. 60. 832-833 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 飯島信司: "Immortalization of human endothelial cells by temperature sensitive simian virus 40" Animal Cell Technology : Developments towards the 21st century. 57-61 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 飯島信司: "Immortalization of human endothelial cells by temperature sensitive simian virus 40" The Sixth Annual Meeting of Japanese Association for Animal Cell Technology(JAACT'93)Program & Abstracts. 46 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 飯島信司: "Isolation of immortal human endothelial cells" Bioscience,Biotechnology,and Biochemistry. 59. 912-914 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 飯島信司: "Blocking adhesion of cancer cells to endothelial cell types by S.agalactiae type-specific polysaccharides" Cytotechnology. 22. 205-210 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 飯島信司: "細菌表面多糖が示すガン細胞接着阻害活性" 化学工学. 60. 832-833 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 飯島 信司: "Immortalization of human endothelial cells by temperature sensitive simian virus 40" Animal Cell Technology : Developments towards the 21st century. 57-61 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 飯島 信司: "Immortalization of human endothelial cells by temperature sensitive simian virus 40" The Sixth Annual Meeting of Japanese Association for Animal Cell Technology (JAACT‘93) Program & Abstracts. 46 (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] 飯島 信司: "Isolation of immortal human endothelial cells" Bioscience,Biotechnology,and Biochemistry. 59. 912-914 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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