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Development of a flow type of current application measuring system armed with specific capture and detectability for heavy metal ions

Research Project

Project/Area Number 16550085
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Analytical chemistry
Research InstitutionKanagawa Institute of Technology

Principal Investigator

SATOH Ikuo  Kanagawa Institute of Technology, Department of Applied Chemistry, Professor, 工学部, 教授 (20148125)

Co-Investigator(Kenkyū-buntansha) IIDA Yasuhiro  Kanagawa Institute of Technology, Department of Applied Chemistry, Assistant Professor, 工学部, 講師 (40329305)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥3,000,000 (Direct Cost: ¥3,000,000)
KeywordsMetalloenzyme / Apoenzyme / Immobilized Enzyme / Heavy Metal Ions / Flow Injection Analysis / Electrolysis / Sensor / Enzymatic Analysis / 酵素カラム / アポ酵素活性化法 / 通電
Research Abstract

In the first year, a chemical method for regenerating the apoenzymes with use of chelating agents was applied to the flow-injection analysis and compared with a method based on electrolysis. Two kind of chelating agents such as pyridine dicarboxylate and EDTA were used for regeneration of the apoenzymes. We found that the former efficiently deactivated alkaline phosphatase from E.coli, and that exposure of the latter to the ALP from calf intestine was effective for gaining the apoenzymes. Detail investigation on the alternate conversion processes between regenerating and reactivating apoenzymes demonstrated that PDC should be adsorbed onto the surface of the enzymes immobilized onto the porous glass beads and thereby, coordination to the active center of the enzyme by the substrates should be prevented. Therefore, the enzymes were apparently inhibited by addition of PDC solution.
In the second year, N-dimethylcarboxylsarcosine (DTCS) as masking agents for cobalt(II) ions was added to the sample metal-solutions and then, injected into the once chelator-exposed column. Thus, it was confirmed that reactivation of the enzyme column attributable to zinc(II) ions could be observed. In conclusion, selective microdetermination of zinc(II) ions could be achieved by using the masking agent. Furthermore, excess amount of zinc(II) ions adsorbed onto the CPG was detected by monitoring the effluents from the enzyme column. Therefore, ALP from E.coli was immobilized onto a net type of gold instead of using CPG as the supporting material and thereby, reproducibility of the assay for the enzyme activity attributable to reactivation of the apoenzyme column could be significantly improved. These results can provide crucial information to microanalysis of heavy metal ions in real samples.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (20 results)

All 2006 2005 2004 Other

All Journal Article (17 results) Book (3 results)

  • [Journal Article] SIA of Zn(II) Ions Using a Column Packed with ALP-immobilized beads2006

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Proceedings of the 39^<th> Chemical Sensor Symposium 22・Sup.A

      Pages: 142-144

    • NAID

      40007380861

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] An ALP-column as a selective recognition element for zinc(II) ions in flow streams2006

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Tech. Digests of the 11^<th> Int. Meet. on Chem. Sensors (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] SIA of Zn(II) ions using a column packed with ALP-immobilized beads2006

    • Author(s)
      Ikuo SATOH
    • Journal Title

      Proceedings of the 41st Chemical Sensor Symposium 22・Supplement A

      Pages: 142-144

    • NAID

      40007380861

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Microdetermination of Zinc(II) ions using an ALP-column2005

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Proceedings of the 39^<th> Chemical Sensor Symposium 21・Sup.A

      Pages: 112-114

    • NAID

      40006743283

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Estimation of Removal and Addition of Cofactors from Alkaline Phosphatase and Application of the Process to Determination of Zinc(II) Ions2005

    • Author(s)
      Yasuhiro Iida
    • Journal Title

      Proceedings of the 39~<th> Chemical Sensor Symposium 21・Sup.A

      Pages: 115-117

    • NAID

      40006743284

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Regeneration of an Apoenzyme Column and its Use of a Recognition Element for Microdetermination of Heavy Metal Ions in Flow Streams2005

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Technical Digests of The 6^<th> East Asia Conference on Chemical Sensors

      Pages: 156-157

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Microdetermination of Zinc(II) Ions Using an ALP Column2005

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Proceedings of The 39^<th> Chemical Sensors 21-Sup.A

      Pages: 112-114

    • NAID

      40006743283

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Estimation and Addition of Cofactors from Alkaline Phosphatase and Application of the Process to Determination of Zinc(II) Ions2005

    • Author(s)
      Yasuhiro Iida
    • Journal Title

      Proceedings of The 39^<th> Chemical Sensors 21-Sup.A

      Pages: 115-117

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Regeneration of an Apoenzyme Column and its Use of a Recognition Element for Microdetrmination of Heavy Metal Ions in Flow Streams2005

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Technical Digests of The 6^<th> East Asia Conference on Chemical Sensors, Guilin, China

      Pages: 156-157

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] SIA of Zn(II) Ions using a Column Packed with ALP-Immobilized Beads2005

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Proceedings of The 41^<st> Chemical Sensors 22-Sup.A

      Pages: 142-144

    • NAID

      40007380861

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Regeneration of an apoenzyme column and its use of a recognition element for microdetermination of heavy metal ions in flow streams2005

    • Author(s)
      Ikuo SATOH
    • Journal Title

      Technical Digest of the 6^<th> East Asia Conference on Chemical Sensors

      Pages: 156-157

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Estimation of removal and addition of cofactors from alkaline phosphatase and application of the process to determination of zinc (II) ions2005

    • Author(s)
      Yasuhiro IIDA
    • Journal Title

      Proceedings of the 39th Chemical Sensor Symposium 21・Supplement A

      Pages: 115-117

    • NAID

      40006743284

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Microdetermination of zinc(II) ions using an ALP-column2005

    • Author(s)
      Ikuo SATOH
    • Journal Title

      Proceedings of the 39th Chemical Sensor Symposium 21・Supplement A

      Pages: 112-114

    • NAID

      40006743283

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Flow Injection Analysis of Zinc(II) Ions with an ALP-Column Based on an Apoenzyme Reactivation Method and Application of the Method to Regulation of the Column Activity2004

    • Author(s)
      Yasuhiro Iida
    • Journal Title

      Chemical Sensor VI : Chemica ; and Biological Sensors and Analytical Methods, The Electrochemical Society, Inc. 10

      Pages: 300-302

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Electrolytic regeneration of apoenzymes from metalloenzymes and its application to microdetermination of heavy metal ions2004

    • Author(s)
      Ikuo SATOH, Masashi SEKINE, Yasuhiro IIDA
    • Journal Title

      Chemical Sensors, Supplement B 20

      Pages: 68-69

    • Related Report
      2004 Annual Research Report
  • [Journal Article] An ALP-Column as a Selective Recognition Element for Zinc(II) Ions in Flow Streams

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Technical Digests of The 11^<th> International Meeting on Chemical Sensors (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Biosensors

    • Author(s)
      Ikuo Satoh
    • Journal Title

      Hand Book of Proteins (Eds., A.Ikai et al.) (Asakura Publishing Co., Ltd.) (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] タンパク質の事典2006

    • Author(s)
      佐藤生男(分担執筆)
    • Publisher
      朝倉書店(株)(印刷中)
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] Chemical Sensors VI : Chemical and Biological Sensors and Analytical Methods2005

    • Author(s)
      Y.Iida(分担執筆)
    • Total Pages
      460
    • Publisher
      The Electrochemical Society Inc.
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] Chemical Sensors VI : Chemical and Biological Sensors and Analytical Methods2004

    • Author(s)
      Y.Iida, M.Sekine, S.Nemoto, I.Satoh
    • Publisher
      The Electrochemical Society, Inc.
    • Related Report
      2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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