고농도 세포배양에서 Bacillus thuringiensis의 세포 성장과 포자 형성 속도

Growth Kinetics and Sporulation of Bacillus thuringiensis in High Cell Density Culture

  • 강병철 (동의대학교 공과대학 화학공학과) ;
  • 장호남 (한국과학기술원 화학공학과 및 생물공정연구센터)
  • 발행 : 2001.02.01

초록

유가식배양과 세라믹 막을 장착한 생물반응기에서 운전인 TCRC 조업을 통해서 Bacillus thuringiensis의 고농도 세포배양을 실시하였다. 유가식 배양에서 B. thuringiensis의 세포 성장은 선형적으로 증가하였고, 이것은 세포성장 모델리의 결과와 잘 맞았다. 낮은 세포 성장속에도 불구하고 유가식 배양동안에 포자형성은 관찰할 수 없었고, 이것은 연속배양의 결과와는 반대였다. 유가식 조업 후에 회분식 배양으로 바꾸면 300 g/L의 포도당 공급 농도를 사용했을 때 2.7$\times$$10^9$ CFU/mL 의 포자농도를 얻었다. 생물반응기내에 세라믹 막을 장착한 TCRC 조업에서 포도당 공급 농도의 영향을 결정하였다. 50 g/L의 포도당 농도를 사용했을 때 TCRC 조업에서 82.5 g-cell/L에 해당하는 최대 세포농도 1.8$\times$$10^{10}$ CFU/mL 를 얻었다. TCRC에서 세포성장은 선형적으로 증가하였고 포도당 농도는 제한되었는데 이것은 세포성장은 모델링의 결과와 잘 일치하였다. 1 g/L의 포도당을 공급한 경우 이외에는 TCRC 조업 동안에 포자형성을 관찰할 수 없었다. 50 g/L의 포도당을 공급한 경우 TCRC조업 후에 회분식 배양으로 전환시키면 1.2$\times$$10^{10}$ CFU/mL 의 포자농도를 얻었고, 이것은 연구된 여러 배양형태 중에 가장 높은 포자농도이다. 이 때 최적의 포도당 공급속도는 0.55 g glucose/h로 공급하였을 때였다.

High cell density culture of Bacillus thuringiensis was conducted in fed-batch culture and TCRC using a bioreactor incorporating ceramic membrane filter. Cell growth of B. thuringiensis in fed-batch culture increased linearly, which was well matched by the results of cell growth modeling. In spite of the slower growth rate during fed-batch culture, no spore formation was observed, which was contrary to the results of continuous culture. Changing culture mode to batch culture after fed-batch operation induced a 2.7$\times$$10^9$ CFU/mL spore concentration using a 300 g/L glucose feed concentration. In TCRC operation incorporating ceramic filter within the bioreactor, the effect of glucose feed concentrations on the cell growth and spore formation of B. thuringiensis was determined. A maximum cell concentration of 1.8$\times$$10^{10}$ CFU/ml, which corresponds to 82.6 g-cell/L, was obtained in the TCRC using a 50 g/L glucose feed concentration. In the TCRC, cell growth increased linearly and glucose concentration was limited, which agreed well with the results of cell growth modeling. No spore formation was observed except when 1 g/L of glucose was fed. Changing to batch culture induced a 1.2$\times$$10^{10}$ CFU/mL of spore concentration, which was the highest spore concentration obtained among the various culture modes examined. The optimal glucose feed rate was found to be 0.55 g-glucose/h.

키워드

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