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DEVELOPMENTS OF ENZYME SYSTEMS TOLERANT TO ARTIFICIAL CONDITIONS AND ANALYSIS OF THEIR FUNCTIONS

Research Project

Project/Area Number 10555288
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 生物・生体工学
Research InstitutionKYOTO INSTITUTE OF TECHNOLOGY

Principal Investigator

KUNUGI Shigeru  Department of Polymer Science and Engineering, Kyoto Institute of Technology, Professor, 繊維学部, 教授 (70111929)

Co-Investigator(Kenkyū-buntansha) KAWAI Shouji  TERAMECS Co., Director (Researcher), 常務取締役(研究職)
HIRAGA Kazumi  Department of Applied Biology, Kyoto Institute of Technology, Instructor, 繊維学部, 助手 (50252549)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1999: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1998: ¥3,700,000 (Direct Cost: ¥3,700,000)
KeywordsTolerant Enzyme / High Pressure / High Temperature / Induction / Artificial Conditions / Screening Procedures / Organic Solvents
Research Abstract

Developments of enzyme systems which can be used in rather hard artificial conditions such as high temperature, high salt conditions, high contents of organic solvents, and high pressure, are necessary to the wider applications of enzymatic processes into chemoenzymatic applications.
In this project two extreme artificial conditions, namely solvent-tolerance and pressure-tolerance, are chosen as the targets and in order to obtain microbilas which produce these tolerant enzymes convenient screening procedures are to be investigated. In addition to this, chemical modification methods and amino-acid replacements are also applied to produce the aimed enzyme systems.
Namely, direct observations of microbilas through micrographic methods under high pressure conditions were made possible by investing pressure micrgoraph cell and a very convenient method of pressure-tolerance screening was proposed by use of a commercially available high-pressure pressing apparatus.
For the chemical modifications, high pressure-induced partial refolding of enzyme proteins was utilized to introduce the midifier reagent into the depth of the protein molecules, which is original to the investigator's laboratory.
The functional analysis of the obtained enzymes under artificial conditions were further performed by use of various spectrophotometric methods, such as 4th derivative absorbance, FTIR, fluorescence, and high pressure electrophoresis, both in situ and ex situ manners.

Report

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

    (20 results)

All Other

All Publications (20 results)

  • [Publications] H. Ikeuchi 他: "Activity and stability of a neutral protease from vibrio Sp. (vimelysin) in a pressure-temperature gradient"European Journal of Biochemistry. (In Press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Kunugi 他: "Pressure-assisted cold-denaturation of carboxypeptidase Y"Bulletin of the Chemical Society Japan. 72(12). 2803-2806 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Kunugi 他: "Single-point amino acid substitutions at the 119th residue of thermolysin and their pressure-induced activation"FEBS Letters. 462(3). 231-235 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Kunugi 他: "Studies on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhabitor and thermolysin"European Journal of Biochemistry. 259. 815-820 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] G.-Q. Tang 他: "Salt Effects on Fluorescence spectal Shifts of DNA-bound Hoechst 33258 and reaction Volumes of the Minor Groove Binding"Bulletin of the Chemical Society Japan. 72(5). 1129-1137 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H. Ikeuchi et al.: "Activity and stability of a neutral protease from Vibrio Sp. (vimelysin) in a pressure-temperature gradient"European Journal of Biochemistry. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Kunugi et al.: "Pressure-assisted cold-denaturation of carboxypeptidase Y"Bulletin of the Chemical Society Japan. 72 (12). 2803-2806 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Kunugi et al.: "Single-point amino acid substitutions at the 119th residue of thermolysin and their pressure-induced activation"FEBS Letters. 462 (3). 231-235 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] S. Kunugi et al.: "Studies on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhibitor and thermolysin"European Journal of Biochemistry. 259. 815-820 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] G. -Q. Tang et al.: "Salt Effects on Fluorescence spectral Shifts of DNA-bound Hoechst 33258 and Reaction Volumes of the Minor Groove Binding"Bulletin of the Chemical Society Japan. 72 (5). 1129-1137 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] G. -Q. Tang et al.: "In vitro increases in plasmid DNA supercoiling by hydrostatic pressure"Biochim. Biophys. Acta. 1443. 364-368 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] H.Ikeuchi 他: "Activity and stability of a neutral protease from Vibrio Sp.(vimelysin) in a pressure-temperature gradient"European Journal of Biochemistry. (in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.Kunugi 他: "Pressure-assisted cold-denaturation of carboxypeptidase Y"Bulletin of the Chemical Society Japan. 72(12). 2803-2806 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.Kunugi 他: "Single-point amino acid substitutions at the 119th residue of thermolysin and their pressure-induced activation"FEBS Letters. 462(3). 231-235 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] S.Kunugi 他: "Studies on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhibitor and thermolysin"European Journal of Biochemistry. 259. 815-820 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] G.-Q.Tang 他: "Salt Effects on Fluorescence spectral Shifts of DNA-bound Hoechst 33258 and Reaction Volumes of the Minor Groove Binding"Bulletin of the Chemical Society Japan. 72(5). 1129-1137 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Guo-Qing Tang: "In vitro Increases in Plasmid DNA Supercoiling by Hydrostatic Pressure" Biochemica et Biophysica Acta. 1443. 364-368 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Guo-Qing Tang: "Study on the Salt Effect on DNA-Hoechst 33258 Interactions by Fluorescence Spectral Shifts and Pressure Dependence of the Binding Equilibrium." Bulletin of the Chemical Society of Japan. 72(in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shigeru Kunugi: "Studies on the formation and stability of a complex between Streptomyces proteinaceous metalloprotease inhibitor and thermolysin" European Journal of Biochemistry. (in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shigeru Kunugi: "Effects of ionic additives and ionic comonomers on the temeprature and pressure responsive behavior of thermoresponsive polymers in aqueous solutions" Langmuir. (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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