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Mechanism of killing bacteria, expression of gene encoded enzyme and its genetics

Research Project

Project/Area Number 14360163
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied animal science
Research InstitutionThe University of Tokyo

Principal Investigator

SAKAI Senkiti  The University of Tokyo, Graduate School of Agricultural and life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (80114487)

Co-Investigator(Kenkyū-buntansha) IMAKAWA Kazuhiko  The University of Tokyo, Graduate School of Agricultural and life Sciences, Associate Professor, 大学院・農学生命科学研究科, 助教授 (00291956)
AOKI Fugaku  The University of Tokyo, Graduate School of Agricultural and life Sciences, Associate Professor, 大学院・新領域創成科学研究科, 助教授 (20175160)
ITO Kikuji  The University of Tokyo, Graduate School of Agricultural and life Sciences, Associate Professor, 大学院・農学生命科学研究科, 助教授 (50100045)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥17,000,000 (Direct Cost: ¥17,000,000)
Fiscal Year 2003: ¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 2002: ¥10,700,000 (Direct Cost: ¥10,700,000)
KeywordsL-amino acid oxidase / H_2O_2 generating enzyme / mastitis / mouse milk / cow milk / killing bacteria / ラクトパーオキシダーゼ / 殺菌作用 / ホルモンによる制御機構 / 過酸化水素感受性細菌 / 発生期酸素感受性細菌 / 過酸化水素
Research Abstract

L-Amino acid oxidase (LAO) and lactoperoxidase (LPO) of mouse milk are separated by gel-filtration chromatography.LAO generates H_2O_2 in the presence of L-amino acids and subsequently, LPO generates O through the degradation of H_2O_2. Milk is capable of producing the bacterial toxic H_2O_2 and O The quantity of H_2O_2 was around 300 μmoles/ml milk. In order to examine whether or not LAO and LPO is responsible for killing bacteria in the mammary gland, bacteria (Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Streptococcus agalactiae) are isolated from bovine mastitis milk and cultured with LAO in the presence and absence of LPO. In the presence of L-amino acids, LAO inhibits the growth of Klebsiella pneumoniae and Escherichia coli, and kills Staphylococcus aureus. Streptococcus agalactiae grows normally in the presence of LAO and does not grow in the presence of LAO and LPO. Thus, we divide them into two types : one being H_2O_2-sensitive and the other being O^-sensitive. It is concluded that H_2O_2 and O^-generated in milk is able to prevent the growth of bacteria in the mouse mammary gland.
Cow milk of Holstein and Japanese Black breeds are collected and separated by gel-filtration chromatography. The H_2O_2 generating enzyme is appeared at the peak fraction of 75, being a molecular weight of about 10 k. The eluting profiles of the two breeds are identical. The quantity.of this enzyme in milk is small at early lactation and at late lactation, but is relatively high at mid-lactation. By the incubation for 2 hr at 37℃, the enzyme generates H_2O_2 about 5-fold as compared to that of the 0 hr-incubation.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Nagaoka K, Sakai A, Nojima Y, Imakawa K, Sakai: "A chemokine, interferon (IFN)-g-inducible protein 10 kDa, is stimulated by IFN-t and recruits immune cells in the ovine endometrium"Biology of Reproduction. 68. 1413-1421 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagaoka K, Nojima H, Watanabe, F, Chang, K, Christenson R: "Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interferon-g-inducible protein 10 kDa (IP-10), during early gestation"The Journal of Biological Chemistry. 278. 29048-29056 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nojima H, Nagaoka K, Suda K: "Increase in DNA methylation down-regulates conceptus interferon-tau gene expression"Molecular Reproduction and Development. In press. (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hara KT, Sakuma Y, Sakai S, Nagata M, Aoki F: "Dynamic changes in the expression of protein tyrosine phosphatases during preimplantation mouse development"Journal of Reproduction and Development. 49. 323-328 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Matuda F, Xu N, Kijima S, Sakai S, Imakawa K.: "Analysis of transcriptional control elements in the 5'-upstream region of ovine Interferon-taw gene using feeder independent caprine trophoblast cell line, HTS1"Placenta. 25. 166-175 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagaoka K, Sakai A, Nojima H, Suda Y, Yokomizo Y, Imakawa K, Sakai S: "A chemokine, interferon (IFN)-g-inducible protein 10 kDa, is stimulated by IFN-t and recruits immune cells in the ovine end ometrium."Biology of Reproduction. 68. 1413-1421 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagaoka K, Nojima H, Watanabe F, Chang KT, Christenson RK.: "Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interferon-g-inducible protein 10 kDa (IP-10), during early gestation"The Journal of Biological Chemistry. 278. 29048-29056 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nojima H, Nagaoka K, Suda Y, Sakai A, Imakawa: "Increase in DNA methylation down-regulates conceptus interferon-tau gene expression"Molecular Reproduction and Development. (in press). (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hara KT, Sakuma Y, Sakai S, Nagata M: "Aoki FDynamic changes in the expression of protein tyrosine phosphatases during preimplantation mouse development"Journal of Reproduction and Development. 49. 323-328 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Matuda F, Xu N, Kijima S, Christensen RK, Sakai S, Imakawa KJ: "Analysis of transcriptional control elements in the 5'-upstream region of ovine Interferon-taw gene using feeder independent caprine trophoblast cell line, HTSI"Placenta. 25. 166-175 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nagaoka K, Sakai A, Nojima H, Suda Y, Yokomizo Y, Imakawa K, Sakai: "A chemokine, interferon (IFN)-g-inducible protein 10 kDa, is stimulated by IFN-t and recruits immune cells in the ovine endometrium."Biology of Reproduction. 68. 1413-1421 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nagaoka K, Nojima H, Watanabe F, Chang KT, Christenson: "Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interferon-g-inducible protein 10 kDa (IP-10), during early gestation"The Journal of Biological Chemistry. 278. 29048-29056 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nojima H, Nagaoka K, Suda Y, Sakai A, Imakawa K: "Increase in DNA methylation down-regulates conceptus interferon-tau gene expression"Molecular Reproduction and Development. (In press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hara KT, Sakuma Y, Sakai S. Nagata M, Aoki F: "Dynamic changes in the expression of protein tyrosine phosphatases during preim plantation mouse development"Journal of Reproduction and Development. 49. 323-328 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kuraishi T, Mizoguchi Y, Sun Y, Aoki F, Sakai S: "The casein mRNA decay changes in parallel with the poly(A) tail length in the mouse Mammary gland"Molecular and Cellular Endocrinology. 190 1. 101-107 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sun Y, Nonobe E, Kobayashi Y, Sakai S: "Characterization and expression of L-amino acid oxidase of mouse milk"Journal of Biological Chemistry. 277.21. 19080-19086 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nafaoka K, Sakai A, Imakawa K, Sakai S: "A chemokine, IEN-gamma-inducible protein 10 kDa (IP-10), is stimulated by IFN-taw and immune cells in the ovine endometrium"Biology of Reproduction. (In press). (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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