Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Construction of Overexpression Vectors and Purification of the Oxygenase Component of Alkylphenol Hydroxylase of Pseudomonas alkylphenolia

Pseudomonas alkylphenolia의 알킬페놀 산화효소의 과발현 벡터 제작 및 단백질 정제

  • Lee, Kyoung (Department of Microbiology and Biomedical Science Institute at CWNU, Changwon National University)
  • 이경 (창원대학교 미생물학과 및 생명의과학연구원)
  • Received : 2013.02.04
  • Accepted : 2013.03.04
  • Published : 2013.03.31
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Abstract

Following construction of expression vectors in Escherichia coli, a new procedure involving two-step column purifications with a Fast Performance Liquid Chromatography System was developed for purification of the oxygenase component of alkylphenol hydroxylase of Pseudomonas alkylphenolia. From 50 g wet cake of recombinant E. coli BL21(DE3)(pJJPMO2) cells, 110 mg of pure protein in a heterodimeric form containing a stoichiometric amount of iron were obtained and it exhibited a specific activity of 147 nmole/min/mg.

본 연구에는 대장균에서의 과발현 벡터 개발과 FPLC를 사용한 2단계 컬럼 정제과정을 통해 Pseudomonas alkylphenolia의 alkylphenol hydroxylase의 oxygenase 단백질을 다량으로 정제하는 방법을 개발하였다. 재조합 Escherichia coli BL21(DE3)(pJJPMO2)의 50 g의 wet cake로부터 110 mg의 heterodimer이며 화학량론적 철을 갖는 순수한 단백질을 정제하였으며 147nmole/min/mg의 비활성을 보였다.

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References

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