KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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High Cell Density Culture of Micro-algal Dunaliella bardawil

미세조류 Dunaliella bardawil의 고농도 세포배양

  • 정욱진 (명지대학교 청정기술원/환경·생물공학과) ;
  • 왕만식 (명지대학교 청정기술원/환경·생물공학과) ;
  • 최승인 (명지대학교 청정기술원/환경·생물공학과) ;
  • 정병철 (명지대학교 생명공학과) ;
  • 김주곤 (명지대학교 생명공학과)
  • Published : 1999.04.01
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Abstract

High cell density cultivation of microalga Dunaliella bardawil using nitrogen fed-batch cultures was studied in batch flask. Optimum environmental conditions include concentrated nutrients except NaCl and carbon sources, carbon sources, pH, light, agitation, nitrate and phosphate ions. Cell growth, consumption rates of nitrate and phosphate ions were monitored. Optimal conditions for higher cell density were found to be(in the range tested): 5 times concentrated media(1 times-10 times concentrated media) pH 8.0 (7.0-9.0) white light(blue and red light) 15mM of nitrate (0.94-15mM) 250mM $NaHCO_3$ and $CO_2$ gas. However, the addition of phosphate ions did not enhance the algal maximum cell density and specific growth rate. Nitrate was found to be effective for the cell growth. The maximum cell density of fed-batch culture using nitrate ions in $8.955{\times}106$cells/ml after 189hr incubation.

본 연구에서는 ($\beta$-carotene 생산균주인 미세조류 Dunaliella bardawil을 사용하여 batch flask에서 미세조류의 고농도세포에 관한 최적배양조건(미량원소, pH, agitation speed, nitrate, phosphate, carbon source)을 확립하고자 하였다. 미량원소는 5X 배지에서 교반하였을 때 비생장속도는 $0.0l3hr^{-l}$와 세포농도는 $4.9{\times}10^6$ cells/mL로서 IX. 3X, lOX 배지에서 배양한 것보다. 약 46%, 18%, 69% 높은 세포수율을 얻었으며 세포배양시 교반한 경우, pH는 80에서 최대 세포농도를 얻었다. 초기 nitrate ($KNO_3$)와 phosphate($KH_2PO_4$)의 영향을 조사한 결과 미세조류 생장에 중요한 영양분으로서 질소원의 주입은 매우 효과적임을 확인하였다. 또힌 탄소원으로서 250mM의 $NaHCO_3$와 $CO_2$ 가스를 동시에 사용한 배양조건이 500mM $NaHCO_3$만을 탄소원으로 사용한 실험에 비하여 32% 증가된 세포농도를 나타내었다. light는 white light의 경우 blue light보다 세포생장에 적합하였다. 질소원을 이용한 유기배양시 2회의 nitrate주입만으로써 배양 198hr에 $8.955{\times}10^6$cell/mL의 고농도의 세포를 얻었다.

Keywords

References

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