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Development of Biocatalytic Functional Column Electrolytic Method for Application to High-Performance Biotransformation and Absolute and Sensitive Analysis.

Research Project

Project/Area Number 10660110
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Bioproduction chemistry/Bioorganic chemistry
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

KANO Kenji  Kyoto Univ., Grad. Sch. Agric., Assoc. Prof., 農学研究科, 助教授 (10152828)

Co-Investigator(Kenkyū-buntansha) IKEDA Tokuji  Kyoto Univ., Grad. Sch. Agric., Prof., 農学研究科, 教授 (40026422)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1998: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsNicotinic Acid / 6-Hydroxynicotinic Acid / Column Electrolysis / Electrochemical Complete Hydroxylation / Nicotinic Dehydrogenase / Absolute and Sensitive Analysis / Biotransformation / Electron Transfer Mediator
Research Abstract

Focusing our attention to a biotransformation system from nicotinic acid (NA) into 6-hydroxynicotinic acid (6HNA) by Pseudomonas fluorescens TN-5, the electron transfer from the cell to column electrodes has been realized using ferricyanide as an exogenous electron acceptor. The hydoxylation is remarkably accelerated using such exogenous electron transfer mediator, in place of the native final electron acceptor OィイD22ィエD2. In addition, the succeeding reaction of 6HNA can be completely suppressed by the use of the exogenous electron transfer mediator.
Based on the results, an electrochemically accelerated and 100% biotransformation system has been realized on our column electrolytic system, where Pseudomonas fluorescens TN-5 cells were immobilized on a column electrode. Such complete biotransformation of NA into 6HNA is the first case. This method would be applied to the production of 6HNA as an important starting material of pesticides.
In addition, this method can be successfully applied to the evaluation of the number of NA dehydrogenase in a single cell, where the enzyme immobilized on the column is reduced with NA and after then the reduced enzyme is electrochemically re-oxidized.
Based on a similar principle, NA is bioelectrochemically oxidized at Pseudomonas fluorescens TN-5 cell-immobilized column electrode. The coulometric analysis has allowed highly sensitive and absolute determination of NA down to 0.2 pmol.

Report

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

    (25 results)

All Other

All Publications (25 results)

  • [Publications] Kenji Kano: "Bioelectrochemical Transformation of Nicotinic Acid into 6-Hydroxynicotinic Acid on Pseudomonas fluorescence TN5-Immobilized Column Electrolytic Flow Systems"J.Mol.Catal.B : Enzymatic. 8(4/6). 265-273 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tokuji Ikeda: "Electrochemical Study of Reversible Hydrogenase Reaction of Desulfovibrio vulgaris Cells with Methyl Viologen as an Electron Carrier"Anal.Chem.. 71(9). 1753-1759 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji Kano: "Surface Characterization and On-Line Activity Measurements of Microorganisms by Capillary Zone Electrophoresis"J.Chromatogr.B. 721(1). 31-37 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji Kano: "Biochemical and Electrochemical Characterization of Quinohemoprotein Amine Dehydrogenase from Paracoccus denitrificans"Biochemistry. 38(21). 6935-6942 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji Kano: "Role of 2-Amino-3-Carboxy-1, 4-Naphthoquinone, a Strong Growth Stimulator for Bifidobacteria, as an Electron Transfer Mediator for NAD(P)^+Regeneration in Bifidobacterium longum"Biochim.Biophys.Acta. 1428(2). 241-250 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji Kano: "Amperometric Determination of NAD(P)H with Peroxidase-Based H_2O_2-S*ng Electrodes and Its Application to Isocitrate Dehydrogenase Activity in Serum"J.Electroanal.Chem.. 478(1). 33-39 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji kano et al.: "Bioelectrochemical Transformation of Nicotinic Acid into 6-Hydroxynicotinic Acid on Pseudomonas fluorescence TN5-Immobilized Columu Electrolytic Flow Systems"J. Mol. Catal. B : Enzymatic. 8(4/6). 265-273 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Tokuji Ikeda et al.: "Electrochemical Study of Reversible Hydrogenase Reaction of Desulfovibrio vulgaris Cells with Methyl Viologen as an Electron Carrier"Anal. Chem.. 71(9). 1753-1759 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji kano et al: "Surface Characterization and On-Line Activity Measurements of Microorganisms by Capillary Zone Electrophoresis"J. Chromatogr. B. 721(1). 31-37 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji kano et al: "Biochemical and Electrochemical Characterization of Quinohemoprotein Amine Dehydrogenase from Paracoccus denitrificans"Biochemistry. 38(21). 6935-6942 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji kano et al: "Role of 2-Amino-3-Carboxy-1, 4-Naphthoquinone, a Strong Growth Stimulator for Bifidobacteria, as an Electron Transfer Mediator for NAD(P)ィイD1+ィエD1 Regeneration in Bifidobacterium longum"Biochim. Biophys. Acta. 1428(2). 241-250 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji Kano et al: "Amperometric Determination of NAD (P) H with Peroxidase-Based HィイD22ィエD2OィイD22ィエD2-Sensing Electrodes and Its Application to Isocitrate Dehydrogenase Activity in Serum"J. Electroanal. Chem.. 478(1). 33-39 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kenji Kano: "Bioelectrochemical Transformation of Nicotinic Acid into 6-Hydroxynicotinid Acid on Pseudomonas fluorescence TN5-Immobilized column Electrolytic Flow Systems"J.Mol.Catal : Enzymatic. 8(4/6). 265-273 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takuji Ikeda: "Electrochemical Study of Reversible Hydrogenase Reaction of Desulfovibrio vulgaris Cells with Methyl Viologen as an Electron Carrier"Anal.Chem.. 71(9). 1753-1759 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kenji Kano: "Surface Characterization and On-Line Activity measurements of Microorganisms by Capillary Zone Electrophoresis"J.Chromatogr.B. 721(1). 31-37 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kenji Kano: "Biochemical and Electrochemical Characterization of Quinohemoprotein Amine Dehydrogenase from paracoccus denitrificans"Biochemistry. 38(21). 6935-6942 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kenji Kano: "Role of 2-Amino-3-Carboxy-1,4-Naphthoquinone,a Strong Growth Stimulator for Bifidobacteria, as an Electron Transfer Mediator for NAD(P)^+Regeneration in Bifidobactenium longum"Biochim.Biophys.Acta. 1428(2). 241-250 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kenji Kano: "Amperometric Detemination of NAD(P)H with Peroxidase-Based H_2O_2-Sensing Electrodes and Its Application to lsocitrate Dehydrogenase Activity in Serum"J.Electroanal.Chem.. 478(1). 33-39 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Masaki Torimura et al.: "Bioelectrochemically Accelerated Microbial Conbersion of Nicotinic Acid to 6-Hydroxynicotinic Acid on Microorganism-lmmobilized Column Electrolytic System" Chemistry Letters. 1998(4). 295-296. (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masaki Torimura et al.: "Continuous-Flow Column Electrolytic Spectroscopy for Two-Step One-Electron Transfer Reactions" Joumal of Electroanalytical Chemistry. 451(1/2). 229-235 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masaki Torimura et al.: "Mediator-Assisted Continuous-Flow Column Electrolytic Spectroelectrochemical Technique for the Measurement of Protein Redox Potentials-Application to Peroxidase" Analytical Chemistry. 70(22). 4690-4695 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masaki Torimura et al.: "Surface Characterization and On-Line Activity Measurements of Microorganisms by Capillary Zone Electrophoresis" Joumal of Chromatography,B. 721(1). 31-37 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kazuyoshi Takagi et al.: "Mediated Bioelectrocatalysis Based on NAD-Related Enzymes with Reversible Characteristics" Joumal of Electroanalytical Chemistry. 445(1/2). 209-217 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Masa-aki Haga et al.: "Spectroelectrochemical Analysis of Intervalence Band in Mixed-Valence Di-and Tetranuclear Ru Complexes by the Flow-Through Method" Inorganic Chemistry. 37(9). 2320-2324 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 伊藤允好 他: "薬学機器分析" 廣川書店, 350 (1998)

    • Related Report
      1998 Annual Research Report

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

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