Erythritol을 이용한 Bifidobacteria의 생존력과 저장안정성 증대를 위한 세포포집

Cell Entrapment for Bifidobacteria to Increase Viability and Preservative Stability using Erythritol

  • 임태빈 (인하대학교 공과대학 생물공학과) ;
  • 백인걸 (인하대학교 공과대학 생물공학과) ;
  • 정찬섭 (인하대학교 공과대학 생물공학과) ;
  • 류지성 (인하대학교 공과대학 생물공학과) ;
  • 지근억 (서울대학교 생활과학대학 식품영양학과) ;
  • 허병기 (인하대학교 공과대학 생물공학과) ;
  • 허태련 (인하대학교 공과대학 생물공학과)
  • 발행 : 2002.12.01

초록

본 연구는 포집된 bifidobacteria의 위산과 담즙산에 대한 bifidobacteria의 생존율을 증가시키는 것을 목적으로 하였다. Bifidobacteria의 포집을 위한 alginate 선별 실험은 유사위액에 3시간 반응 후 생존율을 측정하여 결정하였다. 2.5%의 Jiwon Co.의 alginate로 포집한 비드에서의 생존율이 Sigma Co.의 alginate로 포집한 비드나 2.0%, 3.0%, 3.5%, 4.0%의 농도에 비해 우수하게 나타났다. Bifidobacteria의 생존율을 향상시키기 위해 2.5% alginate와 함께 보호 효과를 가져올 수 있는 11종의 식품 첨가물질을 가지고 실험을 하였다. 이 실험에서는 erythritol-alginate 비드가 다른 첨가물질들 보다 높은 생존율을 보였다. 또한 erythritol의 최적농도 결정을 위한 실험으로 유사위액에 3시간 반응 후 생존율을 측정한 결과 erythritol의 농도는 1.0%일 때 56%로 가장 우수하게 나타났다. 2.5% alginate 와 1.0% erythritol의 농도로 결정지어진 비드의 크기에 따른 영향 실험에서는 2.0 mm 비드가 다른 크기의 비드보다 bifidobacteria의 생존율이 가장 높았다. Erythritol-alginate 비드는 포집하지 않은 bifidobacteria나 alginate만으로 포집한 bifidobacteria보다 유사위액이나 담즙액에서의 생존율이 높았다. 이러한 비드의 저장성은 5% skimmilk 수용액 상에서 10주간 저온저장을 하였을 경우에 $10^{7}$ 이상의 균수를 유지시킬 수 있었다. 결과적으로 erytritol-alginate비드는 bifidobacteria의 생존율과 저장 안정성에 있어서 다른 첨가물을 이용한 세포 포집의 경우에서보다 이용가치가 높다고 할 수 있다.

In this study, we attempted to increase the survivability of bifidobacteria in simulated gastric juices and bile salts after cell entrapment with alginate and various food additives, such as erythritol, isomalt, palatinose, skim milk, xanthan gum, isomalto-oligosaccharide, fructo-oligosaccharide, galacto-oligosaccharide, pectin, and mono-sodium glutamate. Additionaly, the stability of bifidobacteria during storage was investigated by measuring survival rate at different temperatures, i.e. at 4$^{\circ}C$, 25$^{\circ}C$ and -20$^{\circ}C$. Bifidobacteria were immobilized in alginate beads and the survival rate was monitored. It was found that bifidobacieria entrapped with 2.5%, alginate showed the highest survival rate at 12%. After addition of the various protective agents, erythritol(1%) showed the best protective efficiency with a survival rate of 56.0% among the additives tested when exposed to simulated gastric juices for 3 h. Immobilized cells suspended in 5% skim milk and stored at 4$^{\circ}C$ survived significantly more than cells stored at 25$^{\circ}C$ and -20$^{\circ}C$. Consequently, the study shows that the survival rate of bifidobacteria immobilized in combination with 2.5% alginate beads and 1% erythritol may be signifcantly increased in simulated gastric juices and bile salts.

키워드

참고문헌

  1. N. Eur. J. Dairy Sci. v.51 Man and Microflora Klupach,H.J.
  2. Milchwissenschaft v.52 Bifidobacteria: Characteristics and potential for application in femented milk products Shah,N.P.
  3. Bifidoacteria and Their role Rasic,J.L.;J.A.Kumann
  4. Food Tech. v.45 Bifidobacteria: Their potential for use in america, dairy products Hughes,D.B.;D.G.Hoover
  5. Probiotics: the scientific basis Fuller,R.
  6. J. Appl.Bacteriol. v.66 A review: Probiotics in man and animals Fuller,R. https://doi.org/10.1111/j.1365-2672.1989.tb05105.x
  7. Int. J. Food Microbiol. v.61 Survival of bifidobacteria in yogurt and simulated gastric juice following immobilization in gellan-xanthan beads Sun,W.;W.Griffiths https://doi.org/10.1016/S0168-1605(00)00327-5
  8. J. Nutr. Biochem. v.9 Diet and bacterial colonization: role of probiotics and prebiotics Walker,W.A.;L.C.Duffy https://doi.org/10.1016/S0955-2863(98)00058-8
  9. Hyg. I. Abt. Orig. v.A223 Die farkalflora bei menschen: Ⅰ.Mitteliung: Die zusammensetzung der faekalflora der verscheidenen altersgruppen. Zentralbl. Bakteriolk Mitsuoka T.;K.Hayakawa
  10. J. Dairy Sci. v.80 Survival of lactic acid bacteria in a dynamic model of the stomach and small intestine: validation and the effects of bile Marteau,P.;M.Minekus;R.Havenarr;J.H.J.Huis In't Veld https://doi.org/10.3168/jds.S0022-0302(97)76027-2
  11. Can. Inst. Food. Sci. Technol. J. v.22 Survival of microencapsulated Bifidobacterium pseudolongum in simulated gastric and intestinal juices Rao,A.V.;N.Shiwnarain;J.Maharaj https://doi.org/10.1016/S0315-5463(89)70426-0
  12. Biotechnol. Bioeng. v.38 Diffusion of lactose in κ-carrageenan/locust bean gum gel beads with of without entrapped growing lactic acid bacteria Amaud,J.P.;C.Lacroix https://doi.org/10.1002/bit.260380913
  13. Biotechnol. Prog. v.9 Effect of chelatants on gellan gel rheological properties and setting temperature for immobilization of living bifidobacteria Camelin,I;C.Lacroix;C.Paquin;H.Prevost;R.Cachon;C.Divies https://doi.org/10.1021/bp00021a008
  14. Korean J. Dairy Sci. v.18 Effect of rehydration and immobilization in Ca-alginate on the survival of Lactobacillus casei and Bifidobacterium bifidum Kim,I.K.;Y.J.Baek;Y.H.Yoon
  15. Biotechnol. Bioeng. v.26 Diffusion characteristics of substrates in Ca-alginate gel beads Tanaka,H.;M.Matsumura;I.A.Veliky https://doi.org/10.1002/bit.260260111
  16. J. Pharm. Sci. v.70 Microencapsulation of living cells and tissues Lim,F.;R.D.Moss https://doi.org/10.1002/jps.2600700402
  17. Milchwissenschaft v.43 no.10 Continuous prefementation of milk by entrapped yoghurt bacteria. I Development of the process Prevost,H.;C.Divies
  18. Korean J. Biotechnol. Bioeng. v.14 no.4 Development of cell technology for the improvement of bifidobacteria viability Park,H.K.;K.S.Bae;T.R.Heo
  19. Biotechnol. Biopro. Eng. v.6 Effect of skim milk-alginate beads on survival rate of bifidobacteria Yu,W.K.;T.B.Yim;T.R.Heo https://doi.org/10.1007/BF02931959
  20. Korean J. Biotechnol. Bioeng. v.15 Development of the selection technique of entrapment materials for the viability improvement of entrapped bifidobacteria Lee,K.Y.;C.J.Woo;K.S.Bae;T.R.Heo https://doi.org/10.1002/9783527620968.ch
  21. Biotechnol. Prog. v.13 Production of small, monodispersed alginate beads for cell immobilization Seifert,D.B.;J.A.Phillips https://doi.org/10.1021/bp9700723
  22. Gut v.13 Gastric acid barrier to ingested microorganisms in man: Studieds in vivo and in vitro Gianella,R.A.;S.A.Brotiman;N.Zamcheck https://doi.org/10.1136/gut.13.4.251
  23. The property and application of polyols No,B.S.;S.Y.Kim