High Density Cell Culture of Bifidobacterium by Optimization of Medium Composition and Culture Conditions.

배지조성 및 배양환경 최적화에 의한 Bifidobacterium의 고농도 배양

  • Published : 2002.03.01

Abstract

Bifidobacterium strain was isolated from the feces of brast fed infants. The isolated strain was identified as Bifidobacterium longum by 16S rRNA sequence analysis and named as Bifidobacterium SH2. The MRS medium was modified to obtain high density cells of Bifidobacterium SH2. The optimal medium was determined to be 50 g/L lactose, 10 g/L beef extract, 10 g/L peptone, 5 g/L yeast extract, 7 g/L sodium acetate, 2 g/L ammonium citrate, 2 g/L disodium phosphate,1 g/L tween 80, 0.2 g/L MnSO$_4$ and 0.5 g/L L-cysteine. The pH and temperature were optimized as 5.0 and $37^{\circ}C$, respectively. Through out the optimization of medium composition and culture conditions, the dry cell weight and viable cell count were 2.5 times and 1.8 times higer than those in MRS medium, respectively.

본 연구에서 사용된 Bifidobacterium은 모유를 섭취한 유아의 분변으로부터 분리었으며 16s rRNA 염기서열분석 결과 Bifidobacterium longum속으로 동정되어 Bifidobacterium SH2라 명명하였다. Bifidobacterium SH2의 고농도 배양을 위하여 기본 배지로 MRS 배지를 변형하여 최적화한 결과 최적 배지의 조성은 50g/L lactose, 10 g/L beef extract, 10 g/L peptone, 5 g/L yeast extract, 7 g/L sodium acetate, 2 g/L ammonium citrate, 2 g/L disodium phosphat, 1 g/L tween 80, 0.2 g/L $MnSO_4$, 0.5 g/L L-cysteine이었다. 배양조건을 최적화한 결과 pH는 5.0에서, 온도는$ 37^{\circ}C$에서 균체량이 최대로 나타났다. 배지와 배양 조건을 최적화하여 건조 균체량과 생균수가 MRS 배지 보다 각각 2.7배, 1.8배 증가한 결과를 얻었다.

Keywords

References

  1. Anatoly, B. and M. C. Robin. 1989. Biochemistryand Physiology of Bifidobacteria. CRC Press, Inc. Florida. pp. 1-131.
  2. Buchanan, R. E. and N. E. Gibbons. 1974. Bergey's Manual of Determinative Bacteriology, 8 ed. pp. 669-676.
  3. Gilliland, S. E. 1990. Health and nutritional benefits from lactic acid bacteria. Microbiol. Rev. 87: 175-181.
  4. Hughes,D. B. and D. G. Hoover. 1991. Bifidobacteria - Their potential for use in american dairy products. Food Technol. 45: 74-83.
  5. Hirayama, K. and J. Rafter. 2000. The role of probiotic bacteria in cancer prebention. Microbes Infect. 2: 681-686.
  6. Husain,I.,J. A. Poupard,and R. E Norris. 1991.Influenceof nutrition on the morphology of a strain of Bifidobacterium bifidum. J Bacteriol. 111: 841-844.
  7. Kim, T. S., H. H. Hyun, and H. H. Lee. 1995. Fermentation conditions for production of cell mass and comparison of saccharide utilization in Bifidobacterium longum and B. breve. J Microbiol Biotechnol. 5: 285-291.
  8. Lim, K. S., C. S. Huh, and Y. J. Back. 1990. Studies on the growth of Bifidobacterium bifidum HY-8108 in milk . Effect of addition of growth promoting substances and cultivation with other lactic acid bacteria on the growth of Bifidobacterium bifidum HY-8108. Kor. J Dairy Sci. 12: 172-180
  9. Roy, D., E Dussault, and P. Ward. 1990. Growth requirements of Bifidobacterium strains in milk. Milchwissenschafi. 45: 500-513.
  10. Dubois, M., K. A. Gilles, J. K. Hamilton. 1956. Colorimeteric method for determination of sugars and related substance. Anal. Chem. 28: 350-356. https://doi.org/10.1021/ac60111a017
  11. Stanbury P. E, A. Whitaker, and S. J. Hall. 1994. Principles of fermentation technology, pp 147-166. 2nd ed. Butterworth Heinemann, Boston.
  12. Wood, B. J. B., W. H. Holzapfel. 1995. The Lactic acid bacteria(2). The genera of the Lactic Acid Bacteria. Blackik Academic& Professional, London. Chapter 8. pp. 279-306.
  13. 장영효 1992. Bifodobacterium의 성장촉진물질에 관한 연구. 성균관 대학교 석사학위 논문.