미세조류 Dunaliella bardawil의 고농도 세포배양

High Cell Density Culture of Micro-algal Dunaliella bardawil

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

초록

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$$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.

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.

키워드

참고문헌

  1. J Plant Physiol. v.131 Effect of Irradiance and Nutrient Deficiency on the Chemical Composition of Dunaliella bardawil Ben Amotz and Avron(Volvocales, Chlorophyta) Ben-Amotz A.
  2. Tibtech v.5 Microalgae Biotechnology John R. B.;David M. T.;Joseph C. W.
  3. Chemical J. v.8 베타카로틴 수요 급증 : 항암제 등 의약용 확대 김영석
  4. J. Biol. Chem. v.266 no.21 Co-regulation of a Gene Homologous to early Light-induced Genes in Higher Plants and β-carotene Biosynthesis in the Alga Dunlaiella baradawil Lers A.;Haim, L.;Ada Z.
  5. Plant Cell Physiol. v.31 no.5 Effect of Inhibitors on the Formation of Stereoisomer in the Biosynthesis of β-carotene in Dunaliella bardawil Sadaka A.;Avron M.;Ben Amotz A
  6. J. Appl. Phycol. v.7 New mode of Dunaliella Biotechn ology : Two-phase Growth for β-carotene Production Ben-Amotz A.
  7. Plant Physiol. v.108 Isolation and char acteration of a protein Associated with Carotene globules in the Alga Dunaliella bardawil Katz A.;Carlos J.;Pick U.
  8. Plant Physiol. v.93 Photoinduction of Massive β-carotene Accumulation by the alga Dunaliella bardawil Lers A.;Yael B.;Ada Z.
  9. Plant Physiol. v.91 Mode of Action of the Massively Accumulated β-carotene of Dunaliella bardawil in Protecting the Alga against Damage by Excess Irradiation Ben-Amotz A.;Katz A.;Avron M
  10. J. Food Sci. v.22 HPLC Seperation of Cis-Trans Carotene Isomers in Fresh and Processed Fruits and Vegetables Chandler L. A.;Schwartz S. J.
  11. J. Assoc. off. anal. chem. v.74 no.1 Comparison of Liquid Chromatographic Methods for Determination of Cis-Trans Isomers of β-carotene Constance A. O.;Steven J. S.;George L. C.
  12. Experientia v.38 Mass Production of Spirulina Santillan C.
  13. Annu. Rev.Plant Physiol. Plant Mol. Biol. v.40 Salinity Tolerance of Eukaryotic Marine Algae Kirst G. O.
  14. 한국생물공학회지 v.11 no.3 해양 미세조류 Isochrysis galbana P arke 성장에 대한 환경 및 영양조건의 영향 오유관;박성훈
  15. Biotechnol. Bioeng v.38 High-Density Photoautotrophic Algal Cultures : Design, Construction, and Operation of Novel Photobioreactor System Javanmardian M.;Bernhard O. Palsson
  16. Plant Physiol. v.101 Differential Reactivity of β-carotene isomers from Dunaliella bardawil toward Oxygen Radicals Jimenez C.;Uri P.
  17. CO₂and HCO₃Accumulation by Microalgae Moroney V. M.;David H. H.;Toibert N. E.