KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Efficient Purification Of Fused Ferritin[$F_{H}+F_{L}$] using Silica Powder and Gel Filtration Chromatography

실리카 분말과 젤 여과 크로마토그래피를 이용한 효과적인 융합 페리틴의 정제

  • 허윤석 (충남대학교 공과대학 화학공학과) ;
  • 김인호 (충남대학교 공과대학 화학공학과)
  • Published : 2002.08.01
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Abstract

An iron-storage protein, ferritin is a spherical shell consisting of 24 H-and L-chain subunits. Soluble form of fused($F_{H}+F_{L}$ chain) ferritin was separated from disrupted recombinant E. coii cells, followed by silica powder adsorption. Ferritin was recovered from silica-poweder by distilled water, which was applied to gel filtration chromatography(GFC). Collected ferritin fractions from the GFC were assayed via iron-uptake and its molecular weight determined using GF-HPLC. Fused ferritin showed a higher activity than the M- or L- chain ferritin by two times.

수용성 형태로 존재하는 융합 페리틴을 정제함에 있어서 실리카 분말을 이용한 전처리 공정은 전체 정제 공정효율 증가에 기여하였다. 전처리 공정을 통해 정제된 융합 페리틴의 순도를 높이기 위해 젤 여과 크로마토그래피를 통해 보다 정제된 응합 페리틴을 얻을 수 있었다. 이렇게 정제된 융합 페리틴의 철분결합능력을 분석해본결과 320 moles $F_{H}+F_{L}$ mole로 활성이 우수함을 알 수 있었으며, 분자량 분석을 통해 융합페리틴(40 k dalton)은 trimer와 monomer형태로 존재함을 확인할 수 있었다

Keywords

References

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