Analytical Science and Technology (분석과학)

The Korean Society of Analytical Sciences (한국분석과학회)
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Study on the immobilization of plant glutathione S-transferase for development of herbicide detection kit

제초제 검출 키트 개발을 위한 식물 해독효소 고정화 연구

  • Cho, Hyun-Young (Department of Chemistry, College of Natural Sciences, Chung-Ang University) ;
  • Lee, Jin-Joo (Department of Chemistry, College of Natural Sciences, Chung-Ang University) ;
  • Kong, Kwang-Hoon (Department of Chemistry, College of Natural Sciences, Chung-Ang University)
  • 조현영 (중앙대학교 자연과학대학 화학과) ;
  • 이진주 (중앙대학교 자연과학대학 화학과) ;
  • 공광훈 (중앙대학교 자연과학대학 화학과)
  • Received : 2010.02.10
  • Accepted : 2010.02.16
  • Published : 2010.04.25
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Abstract

Glutathione S-transferase is known to play a crucial role in detoxification in many cases. To develop a herbicide detection biosensor, we in this study attempted to immobilize glutathione S-transferase enzyme on solid supports, polystyrene and agarose, and Na-alginate. These matrixes were attractive materials for the construction of biosensors and might also have utility for the production of immobilized enzyme bioreactors. We also compared the activities of glutathione-S-transferase immobilized OsGSTF3 and free OsGSTF3. The specific activity of the free enzyme in solution was 3.3 higher than the immobilized enzyme. These results suggest that 50% of the enzyme was bound with the catalytic site in polystyrene-alkylamine bead and immobilized enzymes showed 80% remaining activity until 3 times reuse.

Glutathione S-transferase는 식물의 해독작용에 중추적인 역할을 하는 화학 효소이다. 본 연구에서는 제초제 검출 키트 개발에 응용을 위하여 식물 해독작용에 중추적인 역할을 하는 glutathione Stransferase의 고정화 방법을 연구하였다. Chloroacetanilide계 제초제에 높은 효소 활성을 보이는 벼 유래 OsGSTF3에 공유결합을 통한 polystyrene-alkylamine 비드와 리간드결합을 통한 agarose-aminoalkyl 비드,포괄법을 통한 Na-alginate 비드를 이용하여 고정화를 실시하였다. 정제된 OsGSTF3 10 mg을 사용하여 고정화 하였을 때 0.62 mg/g 비드로 polystyrene-alkylamine에 가장 효율적으로 고정화 되었다. 고정화 된 OsGSTF3의 효소 활성은 야생형의 30%를 나타내었으며, 재사용에 의한 효소활성 측정시 3회 까지 처음 활성의 80% 이상을 유지하였다.

Keywords

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