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Molecular Dissection and Application of an Enzyme-Inhibitor System with Special Reference to a Proteinase That Specifically Decomposes a Hardly Soluble Protein in Food Seeds.

Research Project

Project/Area Number 10660118
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 食品科学・栄養科学
Research InstitutionThe University of Tokyo

Principal Investigator

ABE Keiko  Graduate School of Agricultural and life Sciences, The University of Tokyo, Professor, 大学院・農学生命科学研究科, 教授 (10151094)

Co-Investigator(Kenkyū-buntansha) MATSUMOTO Ichiro  Graduate School of Agricultural and life Sciences, The University of Tokyo, Assistant Professor, 大学院・農学生命科学研究科, 助手 (00291328)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 1999: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1998: ¥2,300,000 (Direct Cost: ¥2,300,000)
Keywordswheat grains / cysteine proteinases / gliadain / cystatins / flour processing
Research Abstract

A recombinant enzyme was produced from gliadain we found as a new cysteine proteinase in wheat seeds. It actually decomposed, though in vitro, gliadin as a major wheat protein and was found to be involved in the proteolysis of the storage protein during germination. Next, we cloned wheat cystatins by screening with oryzacystatin cDNA as a probe. Three cDNA clones encoding cystatins were thus obtained and it was disclosed that all these cystatins belong to the so-called "phytocystatin" family. In the mean time, we produced a recombinant species of gliadain by fusion with glutathione S-transferase (GST). The resultant GST-progliadain elicited a proteolytic activity under a weakly acidic condition, which was strongly inhibited by a commercially available preparation of egg cystatin as well as by oryzacystatin.
The activity was also inhibited to a similar extent by two wheat cystatins we obtained recombinantly. All these results suggest the possibility that a system of the proteolysis with wheat cysteine proteinases and its regulation with wheat cystatins can be industrially applied to food processing for flour quality improvement.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] N. Kudo: "Crystallization and preliminary X-ray diffraction studies of a rice cysteine proteinase inhibitor, oryzacystatin-I"J. Biochem.. 123. 578-570 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] I. Matsumoto: "Functional expression and enzymatic properties of two Sitophilus zeamais cysteine proteinases showing different autolytic processing profiles in vitro"J. Biochem.. 123. 693-700 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y. Kusakabe: "Identification of two α-subunit species of GTP-binding proteins, Gα15 and Gαq, expressed in rat taste buds"Biochim. Biophys. Acta. 1403. 265-272 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Asano-Miyoshi: "Identification of taste tissue-specific cDNA clones from a subtraction cDNA library of rat circumvallate and foliate papillae"J. Biochem.. 124. 927-933 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Misaka: "A gustatory cyclic nucleotide-gated channel. CNG gust is expressed in the retina"NeduroReport. 10. 743-746 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Yasuoka: "Two subfamilies of olfactory receptor genes in medaka fish, Oyzias latipes: genomic organization and differential expression in olfactory epithelium"J. Biochem.. 126. 866-873 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kudo: "Crystallization and preliminary X-ray diffraction studies of a rice cysteine proteinase inhibitor, oryzacystatin-I."J. Biochem.. 123. 568-570 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] I. Matsumoto: "Functional expression and enzymatic properties of two Sitophilus zeamais cysteine proteinases showing different autolytic processing profiles in vitro."J. Biochem.. 123. 693-700 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y. Kusakabe: "Identification of two α-subunit species of GTP-binding proteins, Gαl5 and Gαq, expressed in rat taste buds."Biochim. Biophys. Acta. 1403. 265-272 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Asano-Miyoshi: "Identification of taste tissue-specific cDNA clones from a subtraction cDNA library of rat circumvallate and foliate papillae."J. Biochem.. 124. 927-933 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] T. Misaka: "A gustatory cyclic nucleotide-gated channel."CNGgust is expressed in the retina. NeuroReport. 10. 743-746 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] A. Yasuoka: "Two subfamilies of olfactory receptor genes in medaka fish, Oyzias latipes : genomic organization and differential expression in olfactory epithelium."J. Biochem.. 126. 866-873 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kudo: "Crystallization and preliminary X-ray diffraction studies of a rice cysteine proteinase inhibitor, oryzacystatin-I."J. Biochem. 123. 568-570 (1998)

    • Related Report
      1999 Annual Research Report
  • [Publications] I. Matsumoto: "Functional expression and enzymatic properties of two Sitophilus zeamais cysteine proteinases showing different autolytic processing profiles in vitro"J. Biochem. 123. 693-700 ((1998))

    • Related Report
      1999 Annual Research Report
  • [Publications] Y. Kusakabe,: "Identification of two α-subunit species of GTP-binding proteins, Gα15 and Gαq, expressed in rat taste buds."Biochim. Biophys. Acta. 1403. 265-272 ((1998))

    • Related Report
      1999 Annual Research Report
  • [Publications] M. Asano-Miyoshi,: "Identification of taste tissue-specific cDNA clones from a subtraction cDNA library of rat circumvallate and foliate papillae."J. Biochem. 124. 927-933 ((1998))

    • Related Report
      1999 Annual Research Report
  • [Publications] T. Misaka,: "A gustatory cyclic nucleotide-gated channel. CNGgust is expressed in the retina."Neuro Report. 10,. 743-746 ((1999))

    • Related Report
      1999 Annual Research Report
  • [Publications] A. Yasuoka,: "Two subfamilies of olfactory receptor genes in medaka fish, Oyzias latipes : genomic organization and differential expression in olfactory epithelium"J. Biochem. 126,. 866-873 ((1999))

    • Related Report
      1999 Annual Research Report
  • [Publications] M.Asano-Miyoshi: "Identification of taste tissue-specific cDNA clones from a subtraction cDNA library of rat circumvallate and foliate papillae." J.Biochem.124. 927-933 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] N.Kudo: "Crystallization and preliminary X-ray diffraction studies of a rice cysteine" J.Biochem.123. 568-570 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Y.Kusakabe: "Identification of two α-subunit species of GTP-binding proteins,Gα15 and Gαq,expressed in rat taste buds." Biochim.Biophys.Acta. 1403. 265-272 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] I.Matsumoto: "Functional expression and enzymatic properties of two Sitophilus zeamais cysteine proteinases showing different autolytic processing profiles in vitro." J.Biochem.123. 693-700 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Asakura: "Oryzasin as an aspartic proteinase occurring in rice seeds:purification,characterization,and application to milk clotting." J.Agric.Food Chem.45. 1070-1075 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] J.Funaki,: "Purification and characterization of a neutral protease that contributes to the unique flavor and texture of tofu-misozuke." J.Food Biochem.21. 191-202 (1997)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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