KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Optimization of Culture Conditions for the Production of Diphtheria Toxin

디프테리아 toxin 생산을 위한 발효조건 최적화

  • Cho, Min (Division of Chemical Engineering and Biotechnology, College of Engineering, Ajou University) ;
  • Ryu, Yeon-Woo (Division of Chemical Engineering and Biotechnology, College of Engineering, Ajou University)
  • 조민 (아주대학교 공과대학 화학생물공학부) ;
  • 유연우 (아주대학교 공과대학 화학생물공학부)
  • Published : 1999.04.01
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Abstract

Experimental studies were carried out to optimize the culture conditions of Corynebacterium diphtheriae for the production of diphtheria toxin. A new media which does not contain any meat digest products was selected. The main ingredient of new medium was enzymatic digests of casein known as NZ-Case. In fermenter experiments, the toxin production was increased with the increase of cell growth. The optimum initial pH of media, air flow rate and agitation speed were 7.0, 0.22, vvm and 400 rpm, respectively. The contents of iron and calcium-phosphate precipitate were important for maximal cell growth and toxin production. The optimum concentration of iron was 0.3 mg/L and calcium-phosphate precipitate could serve in gradual supply of iron to maintain the optimal culture condition which is required for enhanced yield of toxin production. In potency test, the potency of toxoid from fermentor culture was higher than that from static culture. When diphtheria toxin is produced by fermentor culture, it is possible to produce higher levels of toxin and better toxoid quality in terms of safety, yield, productivity and immunity.

디프테리아는 Corynebacterium diphtheriae에 의해 발생되는 호흡기 질병으로 C. diphtheriae의 exo-toxin을 불활성화 시킨 toxoid 백신을 사용하여 예방해 왔다. 현재까지 국내에서는 정치배양 방법으로 디프테리아 toxin을 생산해 왔기 때문에 생산성과 품질에 한계가 있었으며, 이를 극복하기 위해 발효조를 이용한 발효조건 최적화에 대한 연구를 수행하였다. 디프테리아 toxin을 보다 효율적을 얻기 위한 배지로서 beef antigen이 함유되어 있지 않은 casein 유래의 NZ-Case를 선택하였다. 발효조에 의한 toxin 생성은 세포성장과 함께 증가하는 growh-associated form으로 나타났다. 최대의 세포성장과 toxin 생성은 초기 pH가 7.0인 배지에서 0.22vvm의 통기와 400rpm의 교반조건에서 얻을 수 있었다. 또한 최대의 toxin 생성을 위한 최적의 iron 이온의 농도는 0.3mg/L 이었으며, morgamcphospale의 첨가에 의한 calcium-phosphate 침전이 배지 내에 iron 이온의 농도조절을 위하여 요구되었다. 면역원성을 확인하기 위한 역가시험 결과 발효조 배양에서 얻은 toxoid는 4단위에서 모든 guinea-pigs가 생존하여 2단위인 정치배양 toxin에비해 월등히 우수한 것으로 판단되었다. 결국 발효조 배양으로 toxin을 생산할 경우 부작용이 적고 수율과 생산성 및 면역원성 이 우수한 toxid의 생산이 가능하였다.

Keywords

References

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