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Analysis on Ayu. Muscle collagenases induced in the spawning stage

Research Project

Project/Area Number 09660226
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Fisheries chemistry
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

TOYOHARA Haruhiko  KYOTO UNIV.GRADUATE SCHOOL OF AGRIC.ASSOCIATE PROFESSOR, 農学研究科, 助教授 (90183079)

Co-Investigator(Kenkyū-buntansha) KINOSHITA Masato  KYOTO UNIV,GRADUATE SCHOOL OF AGRIC INSTRUCTOR, 農学研究科, 助手 (60263125)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥2,900,000 (Direct Cost: ¥2,900,000)
KeywordsAyu / collagen / collagenase / gelatinase / protease / spawning / matuvation / セラチナーゼ
Research Abstract

Meat texture of ayu deteriorates remarkably in the spawning season. Our recent studies shows that the enzymatic breakdown of collagen in the pericellular connective tissues is responsible for the deterioration, but the protease(s) involved in the breakdown has not yet been identified. Our primary goal in the present study was the identification of the protease(s).
In the first year (1997), we made an attempt to purify the enzymes from cultured ayu muscle by DEAE or CM cellulose chromatography followed by gelatin affinity chromatography which was prepared by ayu type I collagen. By CM cellulose chromatography, we could detect a specific 70kDa activity in the extracts from the muscle of the spawning season in the gelatin zymographic analysis. Further purification of the activity fraction by a gelatin affinity column, 70kDa serine-type activity and 68kDa metallo-type activity were separately eluted in the non-adsorbed and adsorbed fractions, respectively. These results suggested that the 60-70kDa activities usually detected in ayu muscle of the spawning stage are composed of two distinct protease activities of serine-type and metallo-type.
In the second year (1998), we examined the relationship between the activity level of the proteases and the degree of maturation by using ayu caught in the Kuzuryu River. As a result, 68kDa and 62kDa activities were specifically induced in the fish with the gonado-somatic index of 4.7 and 6.0 (male) and 11.1 and 14.3 (female). These results suggested that the protease activities derived purified in the first year actually expressed in the muscle of wild ayu.
Through these studies, we could identify the proteases possibly involved in the deterioration of collagen in the spawning season of ayu. As a next step, it is required to reveal the mechanism of the induction of these activities.

Report

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

    (21 results)

All Other

All Publications (21 results)

  • [Publications] M.TOYOHARA: "Effect of mocturction on the contents of free and bound forms of hydroxy proline in ayu muscle" Fisheries Science. 63. 843-844 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] M.TOYOHARA: "Effect of hormane administration on the breukdown of muscle protein in ayu" Fisheries Science. 64. 419-422 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota: "Occurrence of gelationolytic actiuties in ayu muscle in the spawning stage" Fisheries Science. 64. 439-442 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota: "Gelectinolytic activities in ayu muscle in the spauning stage" Fisheries Science. 64. 1001-1002 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota: "Novel gelatinolytic activities in rat agens" Biochem.Biol.Intern. in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 豊原 治彦: "コラーゲンの代謝-魚類,水産学シリーズ111「魚介類の細胞外マトリックス」" 恒星社厚生閣, 29-37 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S Kubota: "Textual Change by Sexual Maturition, in "Exvacellular matrix of fish and shellfish." Research Signpost in press, (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Toyohara, K.Ito, K.Touhata, M.Kinoshita, S.Kubota, K.Sato, K.Ohtsuki, and M.Sakaguchi: "Effect of maturation on the contents of free and bound forms of hydroxyproline in Ayu muscle" Fisheries Science. 63. 843-844 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Toyohara, K.Ito, K.Touhata, M.Kinoshita, S.Kubota, K.Sato, K.Otsuki, M.Sakaguchi: "Effect of hormone administration on the breakdown of muscle protein in ayu" Fisheries Sci.64. 419-422 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota, H.Toyohara, and M.Sakaguchi: "Occurrence of gelatinolytic activities in yellowtail tissues" Fisheries Sci.64. 439-442 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota, M.Kinoshita, H.Toyohara, and M.Sakaguchi: "Gelatinolytic activities in ayu muscle in the spawning atage." Fisheries Sci.64. 1001-1002 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota, Y.Yokoyama, H.Toyohara, and M.Sakaguchi: "Novel gelatinolytic activities in rat organs." Biochem.Mol.Biol.Intern.(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Toyohara and M.Sakaguchi: "Collagen metabolism-fish" Fisheries Science Series 111 "Extracellular Matrix in Fish and Shellfish" Ed.by S.Kimura, Koseisha-Koseikaku. 29-37 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota, H.Toyohara, and M.Sakaguchi: "Textual Change by Sexual Maturation, in "Extracellular matrix of fish and shellfish"" "Extracellular matrix of fish and shellfish", Research Signpost. (in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Kubota: "Gelutinolytic activities in ayu muscle in the spauming stage" Fisheries Sci.64(6). 1001-1002 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Kubota: "Novel gelatinolytic activities in rat argaus." Biochem.Mol.Biol.Intern.(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Kubota: "Textual change by sexual maturation,in “Extracellular matrix fish and shellfish" Research Signpost(in press), (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] H.Toyohara: "Effect of hovmone administvation on the backdean of muscle protein in ayu" Fisheries Sci.64(3)(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Kubota: "Occurreuce of gelatinolytic activities in yellowtail tissues" Fisheries Sci.64(3)(in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] "Effect of amatuvation on the centents at free and beund forms of hydroxyproline in ayu muscli" Fisheries Sci.63(5). 843-844 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 豊原治彦: "コラーゲンの代謝-魚類、水産学シリーズIII「魚介類の細胞外コトリックス)" 恒星社厚生閣, 97-116 (1997)

    • Related Report
      1997 Annual Research Report

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

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