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Enzyme Chemistry of C-P Compound Synthesis in Rumen Protozoa

Research Project

Project/Area Number 60470141
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 畜産化学
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

HORIGUCHI Masaaki  TOHOKU UNIVERSITY, 農学部, 教授 (20011869)

Co-Investigator(Kenkyū-buntansha) HORIGANE Akira  TOHOKU UNIVERSITY, 農学部, 助手 (10143025)
Project Period (FY) 1985 – 1986
Project Status Completed (Fiscal Year 1986)
Budget Amount *help
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1985: ¥4,800,000 (Direct Cost: ¥4,800,000)
KeywordsC-P compound / Phosphonopyruvate / Rumen protozoa / Tetrahymena / 酵素化学 / ホスホエノールピルビン酸ホスホムターゼ
Research Abstract

1. Some properties and chromatographic behavior of the enzyme which catalyzes formation of 3-phosphonopyruvate (PnPY) in rumen protozoa were studied in an attempt to reveal the role of rumen protozoa in the characteristic metabolism of ruminants.
2. In view of the difficulty of detection and quantitation of free PnPY by HPLC, PnPY-2,4-dinitrophenylhydrazone was separated on a reversed phase column and was detected at 377nm. 0.1 nmol of PnPY could be quantitated.
3. A preliminary research with log phase cells of Tetrahymena, which is a close relative of rumen protozoa, demonstrated that the enzyme localized in the soluble fraction of the cells and required Mg ion and heat stable cofactors. The pH- activity profile of the crude enzyme was bell-shaped and the activity was maximal at about pH8.
4. Mixed protozoa were collected from the contents of sheep rumen, and the cells were fractionated according to the procedures for Tetrahymena. The PnPY-forming enzyme localized in a soluble fraction of the cells, but the activity was very low compared to that in Tetrahymena. On gel permeation chromatography, the enzyme activity in the soluble fraction increased and exceeded the activity in Tetrahymena. The activity was maximal at about pH7.2 and the molecular weight was estimated at about 70k dalton.
5. Results of ion-exchange chromatography revealed that the enzymes of both rumen protozoa and Tetrahymena were basic proteins.
6. Natural occurrence of PnPY has now been established, and all the results were consistent with the biosynthetic mechanism of PnPY through an intramolecular rearrangement of phosphoenolphruvate. However, the final proof for this mechanism of PnPY synthesis must await isolation of the new enzyme, phosphoenolphruvate phosphomutase.

Report

(1 results)
  • 1986 Final Research Report Summary
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] HORIGUCHI,Masaaki;TAKADA,Tatsuyuki: XI Conference on Chemistry and Biochemistry of Natural C-P Compounds.Korea University,Seoul,Korea,November 1,1985.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] TAKADA,Tatsuyuki;HORIGUCHI,Masaaki: XII Conference on Chemistry and Biochemistry of Natural C-P Compound.Shiga University,Otsu,November 14,1986.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] 高田達之,堀口雅昭: 日本農芸化学会昭和62年度大会、講演要旨集. 365 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] 高田達之,鈴木正弘,及川克徳,堀口雅昭: 第78回日本畜産学会大会,1987年.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] M.Horiguchi;T.Takada: "Biosynthesis of C-P Bond in Cell-Free Preparations of Tetrahymena." XI Conference on Chemistry and Biochemistry of Natural C-P Compounds, Korea Univ.,Seoul, Korea, November 1, 1985.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T.Takada;M.Horiguchi: "Purification of C-P Mutase of Tetrahymena." XII Conference on Chemistry and Biochemistry of Natural C-P Compounds, Shiga Univ. Otsu, November 14, 1986.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T.Takada;M.Horiguchi: "Synthesis of 3-Phosphonopyruvate in Cell-Free Preparations of Tetrahymena." Abstr. General Meeting of Agric.Chem.Soc.Jpn. ( 1987 ) p.365.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary
  • [Publications] T.Takada;M.Suzuki;K.Oikawa;M.Horiguchi: "Synthesis of 3-Phosphonopyruvate in Cell-Free Preparations of Rumen Protozoa and Purification of "C-P Mutase"." 78th General Meeting, Jpn.Soc.Zootechn.Sci.,April 2, 1987.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1986 Final Research Report Summary

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Published: 1987-03-31   Modified: 2016-04-21  

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