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CHARACTERIZATION AND APPLICATION OF CHITINOLYTIC ENZYME FROM THE LIVER OF SQUID

Research Project

Project/Area Number 13660208
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Fisheries chemistry
Research InstitutionNihon University

Principal Investigator

MATSUMIYA Masahiro  Nihon University, COLLEGE OF BIORESOURCE SCIENCES, DEPERTMENT OF MARINE SCIENCE AND RESOURCES, ASSOCIATE PROFESSOR, 生物資源科学部, 助教授 (60150702)

Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
KeywordsSQUID / LIVER / CHITINOLYTIC ENZYMES / CHITINASE / CHARACTERIZATION / PURIFICATION / APPRICATION / ACETYLHEXOSAMINIDASE
Research Abstract

1. Characterization was investigated on the 38 kDa and 42 kDa chitinase isozymes from the liver of Japanese common squid. Optimum pH toward colloidal chitin was observed at pH 3.0 for the 38 kDa chitinase, and 3.0 and 9.0 for the 42 kDa chitinase. K_m and k_<cat> of the 38 and 42 kDa chitinases toward a longer substrate, glycol chitin, were 0.071 mg/ml and 1.22/s, and 0.074 mg/ml and 0.196/s, respectively. Both the chitinases decomposed not only chitin but also chitosan (D.A. 95%). Both the chitinases hydrolyzed GlcNAc_n (n=4,5, and 6).
2. Preparation of enzyme solution and some properties of NAHase from the liver of Japanese common squid was investigated. The optimal pH of NAHase was 4.0, and the enzyme activity was stable between pH 4.0 and 6.5. The optimum temperature was 60℃ at 10 min incubation. The enzyme was stable still 60℃ at pH 4.5. The K_m value for pNpGlcNAc was 0.227 mM. Inhibition and/or activation of the enzyme activity by presence of NaCl from 0.1 to 1.0M were not observed.
3. Enzymatic production of N-acetyl-D-glucosamine (GlcNAc) from chitin was investigated using crude enzyme from the liver of Japanese common squid and cuttlefish. The ratio for the activities of chitinase and β-N-acetylhexosaminidase in the crude enzyme was 1:19 for Japanese common squid and 1:20 for cuttlefish. Crude enzyme of Japanese common squid, corresponding to 2 g of liver weight, produced 26.8 mg of reducing sugar from 50 mg of colloidal chitin during 5 days of incubation at 37℃. In the crude enzyme from cuttlefish, 44.4 mg of reducing sugar was obtained under the same reaction conditions. The main product of the produced reducing sugar, analyzed by HPLC, was GlcNAc. These results suggest that squid liver could be a source of chitinolytic enzyme for the enzymatic production of GlcNAc.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Masahiro Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of 38 kDa and 42 kDa chitinase isozymes from the liver of Japanese common squid Todarodes pacificus"Fisheries Science. 68. 603-609 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization and application of chitinolytic enzymes of squid"Fisheries Science Supplement II. 68. 1579-1582 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Purification and some properties of a chitinase isozyme from the liver of Japanese common squid Todarodes pacificus"Fisheries Science. 69. 427-429 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Arakane, Q.Zhu, M.Matsumiya, S.Muthukrishnan, K.J.Kramer: "Properties of catalytic, linker and chitin-binding domains of insect chitinases"Insect Biochemistry and Molecular Biology. 33. 631-648 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of a chitinase from the stomach of chub mackerel"Advances in chitin science. 5. 403-406 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization and application of chitinase isozymes from the liver of Japanese common squid"Advances in chitin science. 6. 265-266 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro Matsumiya, Shuji Karasuda, Kimiko Naruse: "Substrate specificities of chitinases from the stomach of fat greenling Hexagrammos otakii"Advances in chitin science. In press.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro Matsumiya: "Enzymatic production of N-acetyl-D-glucosamine using crude enzyme from the liver of squids"Food Science and Technology Research. In press.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro_Matsumiya.Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of 38 kDa and 42 kDa chitinase isozymes from the liver of Japanese common squid Todarodes pacificus."Fisheries Science. Vol.68. 603-609 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro_Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization and application of chitinolytic enzymes of squid."Fisheries Science Supplement II. Vol.68. 1579-1582 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro_Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Purification and some properties of a chitinase isozyme from the liver of Japanese common squid Todarodes pacificus."Fisheries Science. Vol.69. 427-429 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.Arakane, Q.Zhu, M.Matsumiya.S.Muthukrishnan, K.J.Kramer: "Properties of catalytic, linker and chitin-binding domains of insect chitinases."Insect Biochemistry and Molecular Biology. Vol.33. 631-648 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro_Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of a chitinase from the stomach of chub mackerel."Advances in chitin science. Vol.V. 403-406 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro_Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization and application of chitinase isozymes from the liver of Japanese common squid."Advances in chitin science. Vol.VI. 265-266 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro_Matsumiya, Shuji Karasuda, Kimiko Naruse: "Substrate specificities of chitinases from the stomach of fat greenling Hexagrammos otakii."Advances in chitin science. (In press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro_Matsumiya: "Enzymatic production of N-acetyl-D-glucosamine using crude enzyme from the liver of squids."Food Science and Technology Research. (In press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masahiro Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Purification and some properties of a chitinase isozyme from the liver of Japanese common squid Todarodes pacificus"Fisheries Science. Vol.69 No.2. 427-429 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Y.Arakane, Q.Zhu, M.Matsumiya, S.Muthukrishnan, K.Kramer: "Properties of catalytic, linker and chitin-binding domains of insect chitinase"Insect Biochemistry and Molecular Biology. Vol.33. 631-648 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Masahiro Matsumiya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of a chitinase from the stomach of chub mackerel"Advances in chitin science. Vol.V. 403-406 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 松宮 政弘: "マサバおよびシーラカンス胃のキチン分解酵素について"キチン・キトサン研究. Vol.9, No.2. 162-163 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Purification and some properties of a chitanase isozyme from the liver of Japanese-common squid Todarodes pacifucus"Fisheries Science. 69・2. 427-429 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Y.Arakane, Q.Zhu, M.Matsuimiya, S.Muthukrishnan, K.Kramer: "Properties of catalytic, linker and chitin-binding domains of insect chitinases"Insect Biochemistry and Molecular Biology. 33. 628-631 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of a chitinase from the stomach of chub mackerel"Advances in chitin science. V. 403-406 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization and application of chitinase isozymes from the liver of Japanese common squid"Advances in chitin science. Vol.VI. 265-266 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of 38kDa and 42kDa chitinase isozymes from the liver of Japanese common squid Todarodes pacificus"Fisheries Science. Vol.68, No.3. 603-609 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization and application of chitinolytic enzymes of squid"Fisheries Science. Vo.68 Supplement II. 1579-1582 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 松宮 政弘: "ニュース「第5回アジア太平洋キチン・キトサンシンポジウム」に参加して"キチン・キトサン研究. Vol.8,No.2. 105-107 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Purification and some properties of a chitinase isozyme from the liver of Japanese common squid Todarodes pocificus"Fisheries Science. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization of 38 kDa and 42 kDa chitinase isozymes from the liver of Japanese common squid Todarodes pacificus"Fisheries Science. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] Masahiro Matsumiyhya, Kouji Miyauchi, Atsuishi Mochizuki: "Characterization and application of chitinolytic enzymes of squid"A separate volume of Fisheries Science. (in press).

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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