Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Production of Bacillus anthracis Protective Antigen by Improvement of Culture Condition and Purification Methods

배양조건과 정제방법 개선을 통한 탄저균 방어항원의 생산

  • 김성주 (한양대학교 생화학 및 분자생물학과) ;
  • 조기승 (한양대학교 생화학 및 분자생물학과) ;
  • 최영길 (한양대학교 생명과학과) ;
  • 채영규 (한양대학교 생화학 및 분자생물학과)
  • Published : 2001.03.01
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Abstract

Recently many investigators have devoted considerable attention to the production and purification of PA for antigens, and the preparation of new synthetic medium (RM medium) have solved to increase the yields of the PA but, the low sensitivity of the PA to detect B. anthracis infections has remained as a problem to be solved. This study was undertaken to evaluate the yields of the PA from culture filtrates of B. anthracis Sterne $34F_2$ strain in modified RM medium in which 10 g/l of $NaHCO_3$ and 10g/l of glucose were replaced by 8 g/l and 5 g/l, and the first purification step of PA from culture broth was used hydroxyapatite. The PA was purified by hydroxyapatite column chromatography, DEAE-Sepharose CL-4B column chromatography and Toyo-pearl gel filtration chromatography. The yield of PA from the modified RM medium, 8.6 mg/l.

Bacillus anthracis Sterne 34F$_2$균주로부터 PA를 생산하기 위해 RM배지를 변형하였다. NaHCO$_3$를 8 g/l에서 10 g/l로, glucose를 5 g/l로 첨가하여 새로운 배지조성에서 탄저균을 배양한 후, 배양액을 hydroxyapatite를 이용하여 농축하였다. 농축된 조단백질을 hydroxyapatite column chromatography, DEAE-Sepharose CL-4B column chromatography 및 Toyo-pearl gel filtration chromatography를 사용하여 PA를 정제하였다. 변형된 RM 배지를 사용해 얻은 PA 양은 8.6 mg/l 이었다.

Keywords

References

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