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Basic Studies to Clarify the Role of the β-Primeverosidase in Tea Leaves

Research Project

Project/Area Number 09660112
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Bioproduction chemistry/Bioorganic chemistry
Research InstitutionKYOTO UNIVERSITY (1998-1999)
Shizuoka University (1997)

Principal Investigator

SAKATA Kanzo  Kyoto University, Institute for Chemical Research, Professor, 化学研究科, 教授 (20087563)

Co-Investigator(Kenkyū-buntansha) 坂田 完三  京都大学, 化学研究科, 教授 (20087563)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥1,400,000 (Direct Cost: ¥1,400,000)
KeywordsPrimeverosidase / Glycosidase / Inhibitor / Affinity Choromatography / Disaccharide glycoside / Tea / Amidine / 1-Amino sugar / Camellia sinensis / 茶 / グリコシダーゼ阻害剤 / アミジン / アフィニティー / 配糖体 / β-プリメベロシダーゼ / p-aminophenyl 1-thio-β-primeveroside / グルコシダーゼ
Research Abstract

To investigate the real role of the β-primeverosidase, which was found by us to be concerned with the floral tea aroma formation during the processing of oolong tea and black tea, in tea plants as well as its occurrence among other plants, this research plan was launched. First of all, we tried to prepare affinity adsorbent fortheenzyme to establish a facile purification method for the enzyme. We started with application of the affinity ligands previously reported for affinity chromatography of a few kinds of glycosidases.
1) Preparation of affinity adsorbents with ρ-aminophenyl 1-thio-β-glycosides as ligands and their evaluation for affinity chromatography of glycosidases from tea leaves.
According to the reported procedures two kinds of ρ-nitrophenyl 1-thio-β-glucoside and 1-thio-β-primeveroside were synthesized. They were catalytically reduced and connected to commercially available matrix Formyl-Cellulofine. A crude glycosidase fractions containing the β-primeverosidase and β-glucosi … More dase was treated with each of the affinity adsorbents. Selective adsorption was not observed.
2)Preparation of affinity adsorbents with 1-amino sugars as ligands and their evaluation for affinity chromatography of glycosidases from tea leaves.
1-β-Amino sugars were found to be easily obtained as crystals using methanolic ammonia. They were reacted with succinic anhydride and 3.3-dimethylglutaric acid anhydride to give acyl products. These amino sugars and their acyl derivatives were subjected to the inhibition test for the crude glycosidase from tea leaves. Each amino sugar showed potent inhibitory activity, but their acyl derivatives were not active anymore. They were judged not to be suitable for the ligands for the affinity chromatographic adsorbent.
All our efforts to prepare affinity adsorbents for the primeverosidase based on the previous knowledge have resulted in vain. So we decided to develop a new ligand for our purpose starting with the 1-β-amino sugars which were confirmed to show potent competitive inhibitory activities against glycosidases and to be easily obtained in high yield.
3)Development of a new type of ligand for affinity adsorbents starting with 1-amino sugars.
1-β-Amino sugars of D-glucose, D-galactose and D-xylose were reacted with methyl phenythioacetimidate hydrobromide to yield new amidinium hydrobomide (1,2 and 3) in high yield (83-95%). These compounds showed high competitive inhibition (ki=9.4×10ィイD1-8ィエD1, 2.4×10ィイD1-6ィエD1 and 2.5×10ィイD1-6ィエD1 M) against each corresponding glycosidases. These activities were also very selective. These ligands are very promising ones for the affinity chromatographic adsorbents for glycosidases. Less

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Ijima,Y.et al.,: "Characterization of β-primeverosidase, being concernde with alcoholic aroma formation in tea leaves to be processed into・・・・"J.Agric.Food Chem.. 46. 1712-1718 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 坂田完三、渡辺修司: "植物の香気前駆体としての配糖体の重要性-茶と花のアルコール系香気生成の分子機構"応用糖質科学. 45. 123-129 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 坂田完三: "烏龍茶の香気生成の秘密を探る"化学と生物. 37. 20-27 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sakata,K.etal.: ""Food for Health in the Pacific Rim"Molecalon basis of alcoholic aroma formation during tea processing"Food&Nutorition Press, Inc, Turnbell, Conneticut, USA. 13 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sakata,K..: ""Chemistory of Caffeinated Bevarages, ACS Symposium Siries"β-Primeverosidase concerned with the floral tea aroma・・…"American Chemical Society(in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Ijima, Y., Ogawa, K., Watanabe, N., Usui, Ohnishi-Kameyama, M., Nagata, T. and Sakata, K: "Characterization of β-primeverosidase, being concerned with alcoholic aroma formation in tea leaves to be processed into black tea, and preliminary observations on its substrate specificity."J. Agric. Food Chem. 46(5). 1712-1718 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kanzo Sakata: "Molecular basis of floral aroma formation in oolong tea processing"Kagaku To Seibutu. 37(1). 20-27 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sakata, K., Watanabe, N., and Usui, T: "Molecular basis of alcoholic aroma formation during tea processing"Food for Health in the Pacific Rim, "Proceedings of the 3rd International Conference of Food Science and Technology,(ed.by J. R. Whitaker, et al)., Davis, California, Foods Nutrition Press, Inc., Trumbull, Connecticut. 93-105 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sakata, K.: "β-Primeverosidase concerned with the floral tea aroma formation during processing of oolong tea and black tea.""Chemistry of Caffeinated Beverages. ACS Symposium Series," ed. by Ho, C. and Parliament, T. H., American Chemical Society, Anaheim, CA.. (in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Guo, W., Hiratake, J., Ogawa, K., Yamamoto, M., Ma, S-J., Usui, T. and Sakata, K: "N-β-Glycosylamidines as potent and highly selective inhibitors of β-glycosidases"J. Amer. Chem. Soc. (in preparation).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Sakata,N.Watanabe and T.Usui: "Food for Health in the Pacific Rim"Food & Nutrition Press,Inc.,Trumbell,Conneticat,USA. 93-105 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ijima,Y.,et al: "Characterization of β-primeverosidase,being concerned with alcoholic aroma formation in tea leaves to be processed into black tea,and preliminary observation on its substrate specificity." J.Agric.Food.Chem.46・5. 1712-1718 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sakata,K.,et al: "Molecular basis of alcoholic aroma formation during tea processing." “Food for Health in the Pacific Rim",ed.by J.R.Whitaker,et al. 93-105 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sakata,K.: "β-Primeverosidase concerned with the floral tea aroma formation during processing of oolong tea and black tea." “Chemistry of Caffeinated Beverages.ACS Symposium Series"ed.by Ho,C.,et al.(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ijima,K., Ogawa,K., et al.: "characterization of β-primeverosidase concerned with alcoholic aroma formation intealeaves to be processed to black tea and colong tea and Preliminary observation on……" Journal of Agricultnal Food Chemistry. 46(印刷中). (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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