Activity and Stability of Glucose Oxidase in Molecular Films Assembled Alternately with Polyions

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概要

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• Anionic glucose oxidase(GOD) was assembled alternately with polycations, namely, poly(ethylenimine)(PEI) and poly(dimethyldiallyl-ammonium chloride)(PDDA), in the preparation of molecular films. Enzymatic activity of the films was investigated by sequential redox reaction with glucose, peroxidase(POD) and DA67 dye. The apparent activity was not influenced by substrate diffusion at up to 5 μg of immobilized GOD(at the area of 5 × 5 mm^2 × 2 faces). This is ascribed to the less dense packing of the alternate molecular film compared with Langmuir-Blodgett(LB) films. Immobilized GOD could be released into solution, and its activity was about 80% of native GOD, indicating that the immobilization did not cause significant denaturation. The enzyme activity of the GOD film was maintained for 14 weeks when stored in buffer and in air at 4℃. Activity measurement after incubation at elevated temperatures showed that significant deactivation was not observed up to 50℃. This shows that GOD in the film has higher thermostability than native GOD. The pH profile of the GOD activity in the film became broad and shifted towards higher pH than that of native GOD. The GOD film was also prepared by the premixing method, in which a GOD-polyion complex were assembled alternately with another oppositely-charged polyion. The enzyme activity of the alternate film obtained by premixing was much higher(maximal enhancement, 67-fold) than that of the conventionally assembled films. Better dispersion of GOD in the premixed film appears to enhance the enzyme activity.

• 社団法人日本生物工学会の論文
• 1999-01-25

著者

• Ariga K

  Supermolecules Project Jst(former Jrdc) Kurume Research Park

• Ariga Katsuhiko

  Supermolecules Project Jrdc Kurume Research Park

• Ariga Katsuhiko

  International Center For Materials Nanoarchitectonics National Institute For Materials Science

• Ariga Katsuhiko

  Fuel Cell Materials Center National Institute For Materials Science

• Ariga Katsuhiko

  Advanced Materials Laboratory National Institute For Materials Science

• Ariga Katsuhiko

  Supermolecules Project Jrdc

• Ariga Katsuhiko

  Supermolecules Project Research Development Corporation Of Japan

• Ariga Katsuhiko

  Supermolecules Project Research And Development Corporation Of Japan.

• Ariga Katsuhiko

  Graduate School Of Materials Science Nara Institute Of Science And Technology

• Ariga Katsuhiko

  Department Of Biomolecular Engineering Tokyo Institute Of Technology

• Ariga Katsuhiko

  Graduate School Of Materials Science Nara Institute Of Science And Technology (naist)

• Kimura T

  Fukuoka Industrial Technology Center Biotechnology And Food Research Institute

• 国武 豊喜

  九大工

• 國武 豊喜

  Kyushu Univ. Fukuoka

• Kunitake Toyoki

  Supermolecules Project Jrdc Kurume Research Park

• Kunitake Toyoki

  Department Of Chemical Engineering & Technology Faculty Of Engineering Kyushu University

• Onda Mitsuhiko

  Supermolecules Project Jst(former Jrdc) Kurume Research Park

• Ariga Katsuhiko

  Supermolecules Project Jrdc Kurune Research Center

• Kunitake Toyoki

  Supermolecules Project Jrdc

• Ariga Katsuhiko

  World Premier International (wpi) Research Center For Materials Nanoarchitectonics (mana) National I

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