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Studies of Earthworm Protease for Application of the Catalytic Function

Research Project

Project/Area Number 10556023
Research Category

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

Allocation TypeSingle-year Grants
Section展開研究
Research Field 応用微生物学・応用生物化学
Research InstitutionOkayama Prefectural University

Principal Investigator

NAKAJIMA Nobuyoshi  Okayama Prefectural Univ., Dept.of Nutritional Science Assistant Professor, 保健福祉学部, 専任講師 (10198070)

Co-Investigator(Kenkyū-buntansha) NAKAMURA Kaoru  Kyoto Univ., Institute for Chemical Research Associate Professor, 化学研究所, 助教授 (10101239)
ISHIHARA Kohji  Kyoto Univ.of Education, Dept.of Chemistry Instructor, 教育学部, 助手 (50273537)
SUGIMOTO Manabu  Okayama Univ., Research Institute for Bioresources Instructor, 資源生物科学研究所, 助手 (20216336)
HAMADA Hiroki  Okayama Univ. of Science, Dept.of Applied Science Professor, 理学部, 教授 (10164914)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥7,700,000 (Direct Cost: ¥7,700,000)
Fiscal Year 2000: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1998: ¥3,700,000 (Direct Cost: ¥3,700,000)
KeywordsEarthworm / Fibrinolytic enzyme / Protease / Fibrinolysis / Autolysis / Stability / Gene expression / Nucleotide sequence / Serine protease / fibrirolytic enzyme / Earthworm / Gene cloning / DNA sequence / Lumbricus rubellus / autolysate / Fibrirolytic enzyme / gene cloning / Sequence
Research Abstract

Earthworm serine proteases were investigated for application of the catalytic function and the many useful results were obtained as bellows.
1. Purification and characterization of the proteases.
2. Chemical modification of the protease as a therapeutic enzyme.
3. Construction of an immobilized enzyme bio-reactor.
4. Analysis and utilization of the earthworm autolysate
5. Gene cloning and expression of the proteases.
6. Investigation of structure, function, and stability of the proteases.
7. Investigation of DNA and amino acid sequences of the proteases.
8. Large-scale production of the recombinant protease from Yeast cells.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] N.Nakajima, et.al.,: "Further characterization of Earthworm Serine Protease : Cleavage Specificity Against Peptide Substrates and on Autolysis."Biosci.Biotechnol.Biochem.,. 63(11). 2031-2033 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Nakajima, et.al.,: "Stable Earthworm Serine Protease : Application of the Protease Function and Usefulness of the Earthworm Autolysate."J.Biosci.Bioeng.,. 90(2). 174-179 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Sugimoto and N.Nakajima: "Molecular Cloning, Sequencing, and Expression of cDNA Enco ding Protease having Fibrinolytic Activity from Earthworm."Biosci.Biotechnol.Biochem.,. 65(in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] K.Ishihara,N.Nakajima, et.al.: "A chemoenzymatic Synthesis of Aromatic Carboxylic acid vinyl esters."J.Mol.Catal.B:Enzymatic,. 7. 307-310 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Nakajima, et.al.,: "Degradation of Pectic Substances by Two Pectate Cyases from a Human Intestinal Bacterium, Clostridium butyticum-beijerinckii."J.Biosci.Bioeng.,. 88(3). 331-333 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Nakajima, et.al.,: "Lipase-catalyzed Direct and Regioselective Acylation of Flavonoid Glucoside for Mechanistic Investigation of stable Plant Pigments."J.Biosci.Bioeng.,. 87(1). 105-107 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Nakajima, M.Sugimoto, K.Ishihara, K.Nakamura, and H.Hamada: "Further Characterization of Earthworm Serine Proteases : Cleavage Specificity against Peptide Substrates and on Autolysis."Biosci.Biotechnol.Biochem.. 63(11). 2031-2033 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Nakajima, M.Sugimoto, and K.Ishihara: "Stable Earthworm Serine Proteases : Application of the Protease Function and Usefulness of the Earthworm Autolysate."J.Biosci.Biotech.. 90(2). 174-179 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] M.Sugimoto and N.Nakajima: "Molecular Cloning, Sequencing, and Expression of cDNA Encoding Protease Having Fibrinolytic Activity from Earthworm."Biosci.Biotechnol.Biochem.. 65 (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Nakajima, et.al.,: "Further Characterization of Earthworm Serine Protease : deavage Specificity Agniast peptide Substrates and on Autolysis."Biosci.Biotechnol.Biochem.,. 63(11). 2031-2033 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Nakajima, et.al.,: "Stable Earthworm Serine Protease : Application of the Protease Function and Usefulness of the Earthworm Autolysate."J.Biosci.Bioeng.,. 90(2). 174-179 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] M.Sugimoto and N.Nakajima: "Molecular Cloning, Sequencing, and Expression of cDNA Encoding Protease having Fibrinolytic Activity from Earthworm."Biosci.Biotechnol.Biochem.,. 65(in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] K.Ishihara,N.Nakajima, et.al.,: "A Chemoenzymatic Synthesis of Aromatic Carboxylic Acid Vinyl esters."J.Mol.Catal., B : Enzymatic,. 7. 307-310 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Nakajima, et.al.,: "Degradation of Pectic Substances by two Pectate Lyases from a Human Intestinal Bacterium, Clostridium Butyricom-beijeriakii"J.Biosci.Bioeng.,. 88(3). 331-333 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Nakajima, et.al.,: "Lipase-catalyzed Direct and Regioselective Acylation of Flavonoid Glucoside for Medaristic Investigation of stable Plants Pigments."J.Biosci.Bioeng.,. 87(1). 105-107 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Nakajima,Y.Matsuura,et.al.: "Degradation of Pectic Substances by Two Pectate Lyases from a Human Intestinal Bacterium,Clostridium"J.Biosci.Bioeng.,. 88(3). 331-333 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] N.Nakajima,M.Sugimoto,et.al.: "Further Characterization of Earthworm Serine Protease:Cleavage specificity against Peptide Substrates and on Autolysis"Biosci.Biotech.Biochem.,. 63(11). 2031-2033 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] K.Ishihara,N.Nakajima,et.al.H.Hamada,: "A Chemoenzymatic Synthesis of Aromatic Carboxylic Acid Vinyl Esters"J.Mol.Catal.B:Enzymatic. 7. 307-310 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] T.Matsuda,N.Nakajima,K.Nakamura,et.al.: "Two classes of Enzymes of Opposite Stereochemistry in an Organism"J.Org.Chem.. 65. 157-163 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] K.Ishihara,N.Nakajima,et.al.H,Yamaguchi,: "Sterocontrolled Reduction of α-keto esters with Themophilic Actinomycete,Streptomyces thermocyaneoviolaceus"J.Mol.Catal.B:Enzymatic. 8(in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] K.Ishihara,N.Nakajima,et.al.H,Yamaguchi,: "Asymmetric Reduction of α-keto esters with Thermophilic Actinomycete:Purification and Characterization"J.Mol.Catal.B:Enzymatic. 8(in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] H.Hamada,N.Nakajima et al.: "Highly selective transformation by plant catalysts." J.Mol.Cataly.5. 187-189 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] N.Nakajima,K.Ishihara,H.Hamada,et al.: "Lipase-catalyzed Direct and Regioselective Acylation of Flavonoid Glucoside for Mechanistic Investigation of Stable Plant Pigments" J.Biosci.Biotech.87(1). 106-108 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] N.Nakajima,K.Ishihara,K.Nakamura,et al.: "Multi-enzymatic Glucosylation Using Eucalyptus UDP-Glucosyltransferase Coupled UDP-glucose-Fermentation by Bakers' Yeast." Biosci.Biotech.Biochem.63(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Ishihara,N.Nakajima,H.Hamada,et al.: "A Chemoenzymatic Synthesis of Aromatic Acid Vinyl Esters as an Acyl donor for Lipase-catalyzed Transesterification" Syn.Lett.(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Nakamura,N.Nakajima et al.: "How selectivity in microbial reducation can be controlled" J.Chem.Soc.,Chem.Commun.(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] N.Nakajima,K.Ishihara,Y.Matsuura.,et al.: "Degradation of Pectic Substances in Human Intestine:Contribution and Action Pattern of Pectate Lyase from an Intestinal Bacteriam" Biosci・Biotech.Biochem.63(in press). (1999)

    • Related Report
      1998 Annual Research Report

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

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