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Selection of mastitis-resistant cows using the production of L-amino acid oxidase

Research Project

Project/Area Number 13356006
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
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)
HODATE Kouichi  Kitazato University, School of Veterinary Medicine and Animal Science, Professor, 獣医畜産学部, 教授 (30348594)
AOKI Fugaku  University of Tokyo, Graduate School of Frontier Sciences, Associate Professor, 大学院・新領域創成科学研究科, 助教授 (20175160)
伊藤 喜久治  東京大学, 大学院・農学生命科学研究科, 助教授 (50100045)
沢崎 徹 (澤崎 徹)  東京大学, 大学院・農学生命科学研究科, 教授 (00012047)
甫立 孝一  (独)畜産草地研究所, 生理部, 室長(研究職)
Project Period (FY) 2001 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥49,790,000 (Direct Cost: ¥38,300,000、Indirect Cost: ¥11,490,000)
Fiscal Year 2004: ¥8,450,000 (Direct Cost: ¥6,500,000、Indirect Cost: ¥1,950,000)
Fiscal Year 2003: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
Fiscal Year 2002: ¥14,040,000 (Direct Cost: ¥10,800,000、Indirect Cost: ¥3,240,000)
Fiscal Year 2001: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000)
KeywordsL-amino acid oxidase / lactoperoxidase / antibacterial system / mastistis / mouse milk / cow milk / selection index / 過酸化水素産生能 / 化学発光 / 牛乳酵素 / 過酸化水素発生能 / 過酸化水素 / 化学発光法 / オキシダーゼ / L-アミノ酸オキシダーゼ / 乳蛋白質 / 基質特異性 / ミルク中の遊離アミノ酸 / 細菌毒性
Research Abstract

Characterization of L-amino acid oxidase (LAO) in mouse milk
LAO was purified from mouse milk. LAO reacted with L-amino acids in an apparent order of Phe> Met, Tyr> Cys, Leu> His>> other 11 amino acids and produced hydrogen peroxide (H_2O_2) in a dose-and time-dependent manner. An apparent Km was 1.0μM for Phe, 8.1μM for Leu and 20.2μM for His. LAO in milk had a molecular weight of 113 kDa and was converted to a 60-kDa protein by SDS-PAGE. By ion-exchange chromatography, the LAO activity was detected in three different fractions. LAO consisted of two subunits but is electrically heterogeneous. The amino acid sequences of N- and C-terminals, and lysylendopeptidase-digested fragments were the same to those of mouse milk LAO deduced from cDNA. This is the first demonstration showing that LAO is present in milk.
Mechanism of Bacterial Killing by Mouse Milk
After gel-filtration, we detected lactoperoxidase (LPO) activity. Using LAO, LPO and amino acids, H_2O_2 was maintained between 190 and 23 … More 0 nmol/ml from 1 to 4 h. We cultured Escherichia (E) coli, Klebsiella (K) pneumoniae, Staphylococcus (S) aureus, and Streptococcus (S) agalactiae strains. Bacteria increased with a doubling time of 30-40 min. However, S.agalactiae and S.aureus did not grow ; the number of viable bacteria remained the same at 4 h as at 0 h. Escherichia coli and K.pneumoniae grew with doubling times of 98 and 84 min, respectively, and growth was suppressed. Bacterial growth was suppressed in the LAO/LPO/amino acid system.
Hydrogen peroxide generating substance in cow milk
Cow milk contained a H_2O_2 generating substance. After gel-filtration, the fraction with a molecular weight of 20 kDa had H_2O_2 generating activity, and generated it in a linear manner against the time. A H_2O_2 generating activity increased from early to mid-lactation and decreases at the end of lactation. The activity in mastitis milk was lower than that in normal milk. These results suggest that the H_2O_2 generating substance plays an important role for the activation of the LPO-system which is a potent antibacterial system formed by LPO, H_2O_2, and thiocyanate. Less

Report

(5 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (34 results)

All 2004 2003 2002 Other

All Journal Article (20 results) Publications (14 results)

  • [Journal Article] Localization of Janus kinase 2 to the nuclei of mature oocytes and early clevage stage mouse embryo2004

    • Author(s)
      Ito M, Nakasato M, Suzuki T, Sakai S, Nagata M, Aoki F.
    • Journal Title

      Biol Reprod. 71

      Pages: 89-96

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Increase in DNA methylation down-regulates conceptus interferon-tau gene expression.2004

    • Author(s)
      Nojima H, Nagaoka K, Suda Y, Sakai S, Imakawa K, Christensen RK, Sakai S.
    • Journal Title

      Mol. Reprod. Develop. 67

      Pages: 396-405

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Localization of Janus kinase 2 to the nuclei of mature oocytes and early clevage stage mouse embryo.2004

    • Author(s)
      Ito M, Nakasato M, Suzuki T, Sakai S, Nagata M, Aoki F.
    • Journal Title

      Biol Reprod. 71

      Pages: 89-96

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Increase in DNA methylation down-regulates conceptus interferon-tau gene expression.2004

    • Author(s)
      Nojima H, Nagaoka K, Suda Y, Sakai S, Imakawa K, Christensen RK, Sakai S.
    • Journal Title

      Mol.Reprod.Develop. 67

      Pages: 396-405

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Localization of Janus Kinase 2 to the nuclei of mature oocytes and early cleavage stage mouse embryo2004

    • Author(s)
      Ito M, Nakasato M, Suzuki T, Sakai S, Nagata M, Aoki
    • Journal Title

      Biol Reprod. 71

      Pages: 89-96

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Increase in DNA methylation down-regulates conceptus interferon-tau gene expression2004

    • Author(s)
      Nojima H, Nagaoka K, Suda Y, Sakai S, Imakawa K. 他
    • Journal Title

      Mol.Reprod.Develop. 67

      Pages: 396-405

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Dynamic changes in the expression of protein tyrosine phosphates during pre-implantation mouse development : semi-quantitation by real-time PCR.2003

    • Author(s)
      Hara K, Sakura Y, Sakai S, Nagata M, Aoki F.
    • Journal Title

      J. Reprod. Devel. 49

      Pages: 323-328

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A chemokine, interferon(INF)-γ-inducible protein 10 kDa, is stimulated by IFN-τ and recuits immune cells in the ovine endometorium.2003

    • Author(s)
      Nagaoka K, Sakai A, Nojima H, Suda Y, Yoshioka Y, Imakawa K, Sakai S, Christensen RK.
    • Journal Title

      Biol Reprod. 68

      Pages: 1413-1421

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Involvement of coactivator CBP in the regulation of conceptus IFN-tau gene expression.2003

    • Author(s)
      Xu N, Takahashi Y, Imakawa K, Sakai S, Christensen RK.
    • Journal Title

      Mol. Reprod. Develop. 65

      Pages: 23-29

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interone-□-inducible protein 10 Kda (IP 10), during early gestation.2003

    • Author(s)
      Nagaoka K, Nojima H, Watanabe F, Chang KT, Christensen RK, Sakai S, Imakawa_K.
    • Journal Title

      J. Biol. Chem. 278

      Pages: 29048-29056

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Dynamic changes in the expression of protein tyrosine phosphates during pre-implantation mouse development : semi-quantitation by real-time PCR.2003

    • Author(s)
      Hara K, Sakuma Y, Sakai S, Nagata M, Aoki F.
    • Journal Title

      J.Reprod.Devel. 49

      Pages: 323-328

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Involvement of coactivator CBP in the regulation of conceptus IFN-tau gene expression.2003

    • Author(s)
      Xu N, Takahashi Y, Imakawa K, Sakai S, Christensen RK.
    • Journal Title

      Mol.Reprod.Develop. 65

      Pages: 23-29

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Regulation of blastocyst migration, apposition, and initial adhesion by a chemokine, interone-γ-inducible protein 10 Kda (IP10), during early gestation.2003

    • Author(s)
      Nagaoka K, Nojima H, Watanabe F, Chang KT, Christensen RK, Sakai S, Imakawa K.
    • Journal Title

      J.Biol.Chem. 278

      Pages: 29048-29056

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Regulation of blastosyst migration, apposition and initial adhesion by a chemokine, IFN-gamma inducible protein 10 kD2003

    • Author(s)
      Nagaoka K, Nojima H, Watanabe F, Christenzen RK. 他
    • Journal Title

      J.Biol.Chem. (2003) 278

      Pages: 29048-29056

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Characterization and expression of L-amino acid oxidase of mouse milk.2002

    • Author(s)
      Sun Y, Monobe E, Kobayashi Y, Aoki F, Yamamoto K, Sakai S.
    • Journal Title

      J. Biol. Chem. 277

      Pages: 19080-19086

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The casein mRNA decay changes in parallel with the poly(A) tail length in the mouse mammary gland.2002

    • Author(s)
      Kuraishi T, Mizoguchi Y, Sun Y, Aoki F, Imakawa K, Saki S.
    • Journal Title

      Mol. Cell. Endocrinol 190

      Pages: 101-107

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Involvement of chromatin structure in the regulation of mouse zygotic gene activation.2002

    • Author(s)
      Cho T, Sakai S, Nagata M, Aoki F.
    • Journal Title

      Anim. Sci. J. 73

      Pages: 113-122

    • NAID

      10020368611

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Characterization and expression of L-amino acid oxidase of mouse milk.2002

    • Author(s)
      Sun Y, Monobe E, Kobayashi Y, Aoki F, Yamamoto K, Sakai S.
    • Journal Title

      J.Biol.Chem. 277

      Pages: 19080-19086

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The casein mRNA decay changes in parallel with the poly(A) tail length in the mouse mammary gland.2002

    • Author(s)
      Kuraishi T, Mizoguchi Y, Sun Y, Aoki F, Imakawa K, Saki S.
    • Journal Title

      Mol.Cell.Endocrinol 190

      Pages: 101-107

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Involvement of chromatin structure in the regulation of mouse zygotic gene activation.2002

    • Author(s)
      Cho T, Sakai S., Nagata M, Aoki F.
    • Journal Title

      Anim.Sci.J. 73

      Pages: 113-122

    • NAID

      10020368611

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 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, IFN-gamma-inducible protein 10kDa(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
  • [Publications] Kuraishi, T., Mizoguchi, Y., Sun, Y., Sakai, S 他: "The casein mRNA decay changes in parallel with the polly(A) tail length in the mouse mammary gland"Molecular and Cellular Endocrinology. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Sun, Y., Nonobe, E., Acki, F., Sakai, S 他: "Characterization and expression of L-amino acid oxidase of mouse milk"Journal of Biological Chemistry. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Sun, Y., Kuraishi, T., Aoki, F., Sakai, S: "Hormonal regulation of mitochondrial Tim23 gene expression in the mouse mammary gland"Molecular and Cellular Endocrinology. 172. 177-184 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Yoneda, M., Shiraishi, J., Kuraishi, T., Sakai, S 他: "Gastric proteinase digestion of caseins in new born pups of the mouse"Journal of Dairy Science. 84. 1851-1855 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Fuchimoto D, Mizukoshi A, Sakai S, Aoki, F.: "Posttranscriptional regulation of cyclin A1 and cyclin A2 during mouse oocyte meiotic maturation and preimplantation"Developmental Biol. ogy and Reproduction. 65. 986-993 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hara, K., Sakai S., Nagata M., Aoki F: "Expression of protein tyrosine phosphatases in the preimplantation mouse embryos"Animal Science Journal. 72. 404-409 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 森田琢磨, 酒井仙吉, 唐澤 豊, 近藤誠司: ""家畜"のサイエンス"文永堂出版. 189 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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