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Structure Determination and Development of Purification Process of Taste-modifying protein, Miraculin

Research Project

Project/Area Number 62880021
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionYokohama National University. Faculty of Education

Principal Investigator

KURIHARA Yoshie  Yokohama National Univ. Fac. of Education, Professor, 教育学部, 教授 (90017715)

Co-Investigator(Kenkyū-buntansha) KOIMAI Naoyoshi  Mitajiri Chemical Industries, Ltd. Research and Development, Researcher, 研究員
NAKAYA Kazuyasu  Showa University, School of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助教授 (40053855)
NAKAMURA Yasuharu  Showa University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (70053759)
KURIHARA Kenzo  Hokkaido Univ. Fac. of Pharmaceutical Sciences, Professor, 薬学部, 教授 (00016114)
ASOH Takeo  Yokohama National Univ. Fac. of Education, Professor, 教育学部, 教授 (40017736)
Project Period (FY) 1987 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 1989: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1988: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 1987: ¥5,600,000 (Direct Cost: ¥5,600,000)
KeywordsTaste-modifiers / Taste-modifying protein / Asparagin-linked oligosaccharide / Miraculin / Miracle fruit / Glycoprotein / Anti-miraculin IgG / ミラクリンの糖鎖構造 / ミラクリンのジスルフィド結合 / ミラクルフルーツ / ミラクリンの一次構造
Research Abstract

1. The taste-modifying protein, miraculin, has the unusual property of modifying a sour taste into a sweet taste. We found that miraculin was extracted with 0.5 M NaCi solution. Miraculin was purified from the extracted solution by ammonium sulfate fractionation, CM-Sepharose ion-exchange chromatography, and gel-filtration. The purified miraculin thus obtained gave a single sharp peak in reverse phase high performance liquid chromatography, indicating that it is highly pure.
2. The complete amino acid sequence of miraculin was determined by an automatic Edman degradation method. Miraculin was a single polypeptide with 191 amino acid residues. The calculated molecular weight based on the amino acid sequence and the carbohydrate content was 24,600. The carbohydrate chains were elucidated to link to Asn-42 and Asn-186.
3. The structure of each N-linked oligosaccharides isolated was analyzed. It was found that five oligosaccharides are a series of compounds with xylose-containing common structural core. A variety of oligosaccharide structure are significant for two glycosylation sites, Asn-42 and Asn-186. Two new oligosaccharides with unusual structures containing complex-type were characterized.
4. We have developed an enzyme-linked lmmunoabsorbent assay for miraculin. The amounts of miraculin in miracle fruit increased dramatically seven weeks after pollination and reached a maximum at eight weeks. These results suggest that the biosynthesis of miraculin increases dramatically during the final stage of maturation of miracle fruit.

Report

(3 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] N.Takahashi et al.: "Structural Study of Asparagine-linked Oligosaccharide Moiety of Taste-modifying protein,Miraculin" J.Biol.Chem.265. (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] S.Theerasilp et al.: "Complete Amino Acid Sequence and Structure Characterization of Taste-modifying Protein,Miraculin" J.Biol.Chem.264. 6655-6659 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] M.Meada et al.: "Studies on the Taste Modifiers.II.Purification and Structure Determination of Gymnemic Acid,Antisweet Principle from Gymnema sylvestre Leaves" Tetrahedron Lett.30. 1547-1550 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] S.Theerasilp et al.: "Complete Purification and Characterization of the Taste-modifying Protein,Miraculin,from Miracle Fruit" J.Biol.Chem.263. 11536-11539 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] S.Nakajo et al.: "A Quantitative Enzyme Immunoassy for Miraculin in Miracle Fruit" Chemical Senses. 13. 663-669 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Y.Kurihara et al.: "Studies on the Taste Modifiers.I.Purification and Structure Determination of Sweetness Inhibiting Substance in Leaves of Ziziphus jujuba" Tetrahedron. 44. 61-66 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Y. Kurihara et al.: "Studies on the taste modifiers. I. Purification and structure determination of sweetness inhibiting substance in leaves of Ziziphus jujuba" Tetrahedron. 44. 61-66 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] S. Theerasilp and Y. Kurihara: "Complete purification and characterization of the taste-modifying protein, miraculin, from miracle fruit" J. Biol. Chem.263. 11536-11539 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] S. Nakajo et al.: "A quantitative enzyme immunoassay for miraculin in Richadella dulcifica (miracle fruit)" Chemical Senses. 13. 663-669 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Y. Kurihara: "Purification and structure of taste modifiers: Taste-modifying protein, gymnemic acid and ziziphin" The Beidler Symposium on Taste and Smell, edited by Inglis Miller, Jr. Book Service Association. 75-81 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] M. Maeda et al.: "Studies on the taste modifiers. II. Purification and structure determination of gymnemic acids, antisweet active principle from Gymnema sylvester leaves" Tetrahedron Lett.30. 1547-1550 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] S. Theerasilp et al.: "Complete amino acid sequence and structure characterization of the taste-modifying protein, miraculin" J. Biol. Chem.264. 6655-6659 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] N. Takahashi et al.: "Structural study of asparagine-linked oligosaccharide moiety of taste-modifying protein, miraculin" J. Biol. Chem.265. (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] H. Yamashita et al.: "Purification and complete amino acid sequence of a new type of sweet protein having taste-modifying activity, curculin" J. Biol. Chem.265. (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] N.Takahashi <et>___ー <al>___ー.: "Structual Study of Asparagine-linked Olingosaccharide Moiety of Taste-modifying protein,Miraculin" J.Biol.Chem.(in press). 265. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] S.Theerasilp <et>___ー <al>___ー.: "Complete Amino Acid Sequence and Structure Characterization of Taste-modifying Protein,Miraculin" J.Biol.Chem.264. 6655-6659 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] M.Meada <et>___ー <al>___ー.: "Studies on the Taste Modifiers.II.Purification and Structure Determination of Gymnemic Acid,Antisweet Principle from Gymnema sylvestre Leaves" Tetrahedron Lett.30. 1547-1550 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] S.Theerasilp <et>___ー <al>___ー.: "Complete Purification and Characterization of the Taste-modifying Protein,Miraculin,from Miracle Fruit" J.Biol.Chem.263. 11536-11539 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] S.Nakajo <et>___ー <al>___ー.: "A Quantitative Enzyme Immunoassy for Miraculin in Miracle Fruit" Chemical Senses. 13. 663-669 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] Y.Kurihara <et>___ー <al>___ー.: "Studies on the Taste Modifiers.I.Purification and Structure Determination of Sweetness Inhibiting Substance in Leaves of Ziziphus jujuba" Tetrahedron. 44. 61-66 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] S.Theerasilp et al.: J.Biol.Chem.264. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] S.Theerasilp;Y.Kurihara: J.Biol.Chem.263. 11536-11539 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] S.Nakajo et al.: Chemical Senses. 13. 663-669 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Maeda et al.: Tetrahedron Lett.(1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] Y.Kurihara et al.: Tetrahedron. 44. 61-66 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Y.Kurihara et al.: Annals.of the New York Acad.Sci.510. 747-749 (1987)

    • Related Report
      1988 Annual Research Report
  • [Publications] Edited by lnglis miller: "Beidler Symposium on Taste and Smell" Book Service Associates Winston-Salem,North Carolina, 75-81 (1988)

    • Related Report
      1988 Annual Research Report

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Published: 1987-04-01   Modified: 2016-04-21  

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