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1992 Fiscal Year Final Research Report Summary

Enzymatic synthesis of physiologically functional oligopeptides consisting of essential amino acids

Research Project

Project/Area Number 02453131
Research Category

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

Allocation TypeSingle-year Grants
Research Field 製造化学・食品
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

MATSUNO Ryuichi  Kyoto Univ., Dept. of Food Sci. & Technol., Professor, 農学部, 教授 (30032931)

Co-Investigator(Kenkyū-buntansha) NAKANISHI Kazuhiro  Okayama Univ., Dept. of Biotechnol., Professor, 工学部, 教授 (90026584)
KIMURA Yukitaka  Kyoto Univ., Dept. of Food Sci. & Technol., Instructor, 農学部, 助手 (70211878)
ADACHI Shuji  Kyoto Univ., Dept. of Food Sci. & Technol., Associate Professor, 農学部, 助教授 (90115783)
Project Period (FY) 1990 – 1992
Keywordsoligopeptide / peptide synthesis / enzymatic synthesis / immobilized enzyme / essential amino acids / Leu-enkephalin / biphasic reaction / basic amino acid
Research Abstract

Oligopeptides possess some physiologically superior functionalities such as higher assimilation rate at the brush boader membrane of the intestine, compared with their constituent amino acids. In this study,enzymatic syntheses of oligopeptides including of essential amino acids have been successfully realized. Thermolysin (TL), alpha-chymotrypsin (CT), and papain (PP) were used in organic solvent or in organic/aqueous biphasic system to synthesize di- and tripeptides by their condensation or transfer action. An example of tripeptide synthesized is Z-GFLNH_2. The tripeptide was continuously synthesized by using two columns packed with immobilized TL and CT, which were connected in series, at an yield of more than 80% for 220 h. The oligopeptides synthesized, which included essential amino acids and histidine, were as follows: Z-K(Z)IOMe (TL;condensation, ca. 100%), Z-TWOEt (PP; transfer, 73%), Z-TWVNH_2 (CT; condensation, ca. 100%), Z-LFOMe (TL; condensation, ca. 100%), Z-MLFOMe (TL; co … More ndensation, ca. 100%), Boc-H(Tos)VOEt (PP; condensation, 91%). The water content in the reaction system, in some cases, significantly affected the mass transfer rates of substrates into immobilized enzymes. An increase of the content resulted in a large mass transfer resistance. We proposed a novel enzymatic method to synthesize the precursor of an opioid peptide, Leu-enkephalin, Z-YGGFLOEt from its constituent amino acid derivatives by using CT, TL, and PP. The validity of the method was demonstrated experimentally. Some peptides including a basic essential amino acid, K were synthesized by trypsin in a mixture of acetonitrile and buffer solution. Examinations were focused on the synthesis of di- or tripeptides consisting of only K. We found the conditions where a high yield of objective could be realized and maintained for a long period. This study showed that a variety of oligopeptides could be synthesized in high yields using only four proteinases, and constructed the system to produce many kinds of products in a relatively small amount, which is directed in bioprocess development. Less

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Yukitaka Kimura: "Continuous synthesis of a tripeptide by successive condensation and transesterification catalyzed by two immobilized proteinases in organic solvent." Agric.Biol.Chem.54. 1433-1440 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kazuhiro Nakanishi: "Long-term continuous synthesis of aspartame precursor in a column reactor with immobilized thermolysin." Appl.Microbiol.Biotechnol. 32. 633-636 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yukitaka Kimura: "Snthesis of peptides consisting of essential amino acids by a reactor system using three proteinases and an organic solvent." Agric Biol.Chem.54. 3331-3333 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yukitaka Kimura: "Enzymatic synthesis of the precursor of Leu-enkephalin in waterimmiscible organic solvent systems." Enzyme Microb.Technol.12. 272-280 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 木村 幸敬: "プロテアーゼを用いる有機溶媒中でのペプチド合成" 日本農芸化学会誌. 65. 1111-1114 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ryuichi Matsuno: "Factors affecting performance of a proteinase-catalyzed bioreactors for oligopeptide syntheses." Ann.N.Y.Acad.Sci.672. 363-373 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yukitaka Kimura: "Enzymatic synthesis of peptides with hydrophilic amino acid in water-miscible organic solvent." Ann.N.Y.Acad.Sci.672. 458-461 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 松野 隆一: "バイオリアクター入門(生物化学実験法28)" 学会出研センター, 63 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yukitaka Kimura, Tomoko Yoshida, Koji Muraya, Kazuhiro Nakanishi, and Ryuichi Matsuno: "Continuous synthesis of a tripeptide by successive condensation and transesterification catalyzed two immobilized proteinases in organic solvent." Agric. Biol. Chem.54. 1433-1440 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kazuhiro Nakanishi, Akira Takeuchi, and Ryuichi Matsuno: "Long-term continuous synthesis of aspartame precursor in a column reactor with an immobilized thermolysis." Appl. Microbiol. Biotechnol.32. 633-636 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yukitaka Kimura, Koji Muraya, Yoichi Araki, Hiroyoshi Matsuoka, Kazuhiro Nakanishi, and Ryuichi Matsuno: "Synthesis of peptides consisting of essential amino acids by a reactor system using three proteinases and an organic solvent." Agric. Biol. Chem.54. 3331-3333 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yukitaka Kimura, Kazuhiro Nakanishi, and Ryuichi Matsuno: "Enzymatic synthesis of the precursor of Leu-enkephalin in water-immiscible organic solvent systems." Enzyme Microb. Technol.12. 272-280 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yukitaka Kimura and Ryuichi Matsuno: "Peptide synthesis in organic solvent by proteinases." Nippon Nogeikagaku Kaishi. 65. 1111-1114 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ryuichi Matsuno, Yukitaka Kimura, Shuji Adachi, and Kazuhiro Nakanishi: "Factors affecting performance of a proteinase-catalyzed bioreactors for oligopeptide syntheses." Ann. N. Y. Acad. Sci.672. 363-373 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yukitaka Kimura, Yoshiko Tari, Shuji Adachi, and Ryuichi Matsuno: "Enzymatic synthesis of peptides with hydrophilic amino acid in water-miscible organic solvent." Ann. N. Y. Acad. Sci.672. 458-461 (1992)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1994-03-24  

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