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Experimental device designed for assay and purification of oxygen-sensitive enzymes and its application.

Research Project

Project/Area Number 02557070
Research Category

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

Allocation TypeSingle-year Grants
Research Field Functional basic dentistry
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

YAMADA Tadashi  Tohoku Univ., School of Dentistry, Prof., 歯学部, 教授 (50005021)

Co-Investigator(Kenkyū-buntansha) TAKAHASHI Nobuhiro  Tohoku Univ., School of Dentistry, Assistant Prof., 歯学部, 助手 (60183852)
IWAMI Yoshimiti  Tohoku Univ., School of Dentistry, Assistant Prof., 歯学部, 助手 (60005030)
ABBE Kazuhiko  Tohoku Univ., School of Dentistry, Associate Prof., 歯学部, 助教授 (40151089)
Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥12,200,000 (Direct Cost: ¥12,200,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1990: ¥10,400,000 (Direct Cost: ¥10,400,000)
KeywordsOxygen sensitive enzyme / Pyruvate formate-lyase / Strictly anaerobic conditions / Oral streptococci / Sugar metabolism / Anaerobic glove box / Enzyme assay / Actinomyces / 酸素活性測定 / 酵素の型変換 / 乳酸脱水素酵素 / レンサ球菌 / ピルビン酸ギ酸リア-ゼ / pHの影響 / 嫌気グロ-ブボックス / 嫌気糖代謝
Research Abstract

An anaerobic glove box, equipped with a purification apparatus and a spectorophotometer, was manufactured as a trial and employed. Several interesting finding were turned out as follows.
Pyruvate formate-lyase (PFL) plays an important role in anaerobic sugar metabolism in oral streptococci. PFL was extremely sensitive to oxygen, and assumed to contain a protein radical near a catalytic site. The reaction of molecular oxygen with the radical seemed to generate further harmful active oxygen and bring about irreversible inactivation of the enzyme rapidly. Reversible inactive form of the enzyme was oxygen torelant since it did not contain the radical, and could be activated by PFL-activating enzyme. Partially purified PFL activating enzyme was quite unstable, but incubation of the enzyme with ferrous ion, selenium ion, pterin, restored a part of the activity. The enzyme seemed to contain "a pterin like cofactor" which was assumed to be exist in xanthine oxidase. The activating enzyme of Streptococcus mutans was triggered by pyruvate, but not oxamate, and quite unstable. The properties of the streptococcal enzyme is considered different from that of Escherichia coli.
It is essential for plaque microorganisms to keep intracellular NADH/NAD balance under anaerobic conditions. Oral streptococci employed "the PFL system" to keep the balance. Lactobacilli metabolized acetate into ethanol to oxidize NADH to keep it. Actinomyces could regenerate NAD via succinate pathway when bicarbonate was available, and increased the rate of acid production.

Report

(4 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (28 results)

All Other

All Publications (28 results)

  • [Publications] K.ABBE: "Oxygen and the sugar metabolism in oral streptococci." Proc.Finn.Dent.Soc.87. 477-487 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N.TAKAHASHI: "Metabolism of intracellular polysaccharide in the cells of Streptococcus mutans under strictly anaerobic conditions." Oral Micobiol.Immunol.6. 299-304 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.ABBE: "Difference in conversion of pyruvate formate-lyase between oral streptococci." J.Dent.Res.70. 551 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.YAMADA: "Sugar metabolism of streptococci in dental plaque and in a test tube." J.Dent.Res.70. 650 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y.IWAMI: "Acid productionby streptococci growing at low pH in a chemostat under anaerobic conditions." Oral Microbiol.Immunol.7. 304-308 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N.TAKAHASHI: "Stimulatory effect of bicarbonate on the glycolysis of Actinomyces viscosus and its biochemical mechanism." Oral Microbiol.Immunol.7. 165-170 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K. Abbe: "Oxygen and the sugar metabolism in oral streptococci." Prof. Finn. Dent. Soc.87. 477-487 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N. Takahashi: "Metabolism of intracellular polysaccharide in the cells of Streptococcus mutans under strictly anaerobic conditions." Oral Micobiol. Immunol.6. 299-304 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K. Abbe: "Difference in conversion of pyruvate formate-lyase between oral streptococci." J. Dent. Res.70. 551 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Yamada: "Sugar metabolism of streptococci in dental plaque and in a test tube." J. Dent. Res.70. 650 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y. Iwami: "Acid production by streptococci growing at low pH in a chemostat under anaerobic conditions." Oral Microbiol. Immunol.7. 304-308 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] N. Takahashi: "Stimulatory effect of bicarbonate on the glycolysis of Actinomyces viscosus and its biochemical mechanism." Oral Microbiol. Immunol.7. 165-170 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y.Iwami: "Acid productionby streptococci grouing at low pH in a chemostat under anaerobic conditions." Oral Microbiol.Immunol.7. 304-308 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] N.Takahashi: "Stimulatory effect of bicarbonate on the glycolysis of Actinomyees viscosus and its biochemical mechanism." Oral.Microbiol.Immunol.7. 165-170 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 山田 正: "「むし歯にならない」との表示はどのように決められるべきか-食品の齲蝕誘発性の検定基準の設定に関する国際的動向-" 歯界展望. 77(5,6). 1109-1119 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 山田 正: "虫歯と砂糖と甘味料" Pal Sweet Diet Report. 20. 1-3 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] N.Takahashi: "Metabolism of intracellular polysaccharide in the cells of Streptococcus mutans under strictly anaerobic conditions." Oral Microbiology and Immunology. 6. 299-304 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] K.Abbe: "Oxygen and the Sugar Metabolism in Oral Streptococci." Proceedindg of Finnish Dental Society. 87. 477-487 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] N.Takahashi: "Stimulatory Effect of Bicarbonate on the Glycolysis of Actinomyces viscosus and its biochemical mechanism." Oral Microbiology and Immunology. 7. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Y.IWAMI: "Acid Production by Streptococci Growing at Low pH in a Chemostat under Anaerobic Conditions." Jounal of Dental Research. 71. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Hata: "Biochemical mechanisms of enhanced inhibition of fluoride on the anaerobic sugar metabolism by Streptococcus sanguis." Journal of Dental Research,. 69. 1244-1247 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] N.Takahashi: "Metabolism of intracellular polysaccharide in the cells of <Streptococcus mutans>___ー under strictly anaerobic conditions." Oral Microbiology and Immunology. 6. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] N.Takahashi: "Stimulatory effect of bicarbonate on the glycolysis of <Actinomyces viscosus>___ー and its biochemical mechanism." Oral Microbiology and Immunology.

    • Related Report
      1990 Annual Research Report
  • [Publications] K.Abbe: "NADH oxidase and Cu^<2+>-mediated oxidative inactivation of Streptococcus salivarius fructosyltransferase." Life Science Advances.(1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Y.Iwami: "Increased lactate production by <Streptococcus mutans>___ー NCIB 11723 at low pH level under anaerobic conditions." Oral Microbiology and Immunology.

    • Related Report
      1990 Annual Research Report
  • [Publications] K.Abbe: "In vitro interconversion of the pyruvate formateーlyase of Streptococcus mutans between the oxygenーsensitive active form and the oxygenーinsensitive inactive form." Oral Microbiology and Immunology.

    • Related Report
      1990 Annual Research Report
  • [Publications] 須賀 昭一: "図説齲蝕学" 医歯薬出版株式会社, 310 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 山田 正: "歯科栄養指導大要" クインテッセンス出版, 215 (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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