Effect of Additives on the Viability of Bifidobacteria Loaded in Alginate Poly-l-lysine Microparticles during the Freeze-drying Process

  • Cui, Jing-Hao (National Research Laboratory for Bioavailability Control, College of Pharmacy, Kangwon National University,College of Pharmacy, Soochow University) ;
  • Cao, Qing-Ri (National Research Laboratory for Bioavailability Control, College of Pharmacy, Kangwon National University) ;
  • Choi, Yun-Jaie (College of Agricultural Life Science, Seoul National University) ;
  • Lee, Kyung-Hoon (Department of Molecular Cell Biology, School of Medicine, Sungkyunkwan University, Clinical Research Institute, Samsung Medical Center) ;
  • Lee, Beom-Jin (National Research Laboratory for Bioavailability Control, College of Pharmacy, Kangwon National University)
  • Published : 2006.08.01

Abstract

Bifidobacteria-loaded alginate poly-l-lysine microparticles (bap microparticles) were prepared using an air atomization method and then freeze-dried. The viability of the bap microparticles was investigated as a function of the amount of the bifidobacteria cultures, and the addition of a yeast extract, cryoprotectants, antioxidants and neutralizer. The size of the bap microparticles with and without the bifidobacteria was $84.8{\pm}28.5\;{\mu}m$ ($mean{\pm}standard$ deviation) and $113.1{\pm}38.5\;{\mu}m$, respectively. The surface morphology was slightly ellipsoid and wrinkled regardless of the incorporating bifidobacteria. The viability gradually decreased with increasing freeze-drying time. Free-flowing powdered bap microparticles were obtained at least 12 h after freeze-drying the wetted slurry of bap microparticles. However, the particles tended to aggregate when either lactose or ascorbic acid was added. The addition of a yeast extract, cryoprotectants (glycerol and lactose), antioxidants ($NaHSO_3$ and ascorbic acid) and neutralizer $(Mg_3(PO_4)_2)$ resulted in a significantly higher viability of the bifidobacteria in the bap microparticles after freeze-drying (0.34-1.84 log) compared with the culture alone.

Keywords

References

  1. Carpenter, J. F., Arakawa, T., and Crowe, J. H., Interaction of stabilizing additives with proteins during freeze-thawing and freeze-drying, Dev. Biol. Stand., 74, 225-239 (1991)
  2. Castro, H. P., Teixeira, P. M., and Kirby, R., Evidence of cell membrane damage in L. bulgarisus following freeze-drying process, J. Appl. Microbiol., 82, 87-94 (1997) https://doi.org/10.1111/j.1365-2672.1997.tb03301.x
  3. Cui, J.-H., Goh, J.-S., Park, S.-Y, Kim, P.-H., and Lee, B.-J., Preparation and physical characterization of alginate microparticles using air atomization method. Drug Dev. Ind. Pharm., 27(4), 309-319 (2001) https://doi.org/10.1081/DDC-100103730
  4. Cui, J.-H., Koh, J.-S., Kim, P.-H., Choi, S.-H., and Lee, B.-J., Survival and stability of bifidobacteria-loaded alginate poly-llysine microparticles. Int. J. Pharm., 210, 51-59 (2000a) https://doi.org/10.1016/S0378-5173(00)00560-3
  5. Cui, J.-H., Shim, J.-M., Lee, J.-S., and Lee, B.-J., Gastric acid resistance of lactobacilli and bifidobacteria in commercial drink and liquid yogurts. Kor. J. Microbiol., 36(2), 161-165 (2000b)
  6. Dave, R. I. and Shah, N. P., Effect of cysteine on the viability of yoghurt and probiotic bacteria in yoghurts made with commercial starter cultures, Int. Dairy J., 7, 537-545 (1997) https://doi.org/10.1016/S0958-6946(97)00053-8
  7. de Valdez, G. F., de Giori, G. S., de Ruiz Holgado, A. P., and Oliver, G., Effect of drying medium on residual moisture content and viability of freeze-dried lactic acid bacteria, Appl. Environ. Microbiol., 49, 413-415 (1985a)
  8. de Valdez, G. F., de Giori, G. S., de Ruiz Holgado, A. P., and Oliver, G., Effect of the rehydration medium on the recovery of freeze-dried lactic acid bacteria, Appl. Environ. Microbiol., 50, 1339-1341 (1985b)
  9. Karney L., Upton, M., and McLoughlin, A., Enhancing the viability of Lactobacillus plantum inoculum by immobilizing the cells in calcium-alginate beads incorporating cryoprotectants, Appl. Environ. Microbiol., 56, 3112-3116 (1990)
  10. Kim, K. I., Baek, Y. J., and Yoon, Y. H., Effect of rehydration media and immobilization in Ca-alginate on the survival of Lactobacillus casei and Bifidobacterium bifidum, Korean J. Dairy Sci., 18, 193-198 (1996)
  11. Lee, B.-J., Cui, J.-H., Park, O.-S., Goh, J.-S., Ahn, T.-S., and Park, S.-Y., Stability and gastric acid resistance of lactobacilli and bifidobacteria in commercial yogurts. Kor. J. Microbiol., 35, 89-93 (1999)
  12. Lee, K.-Y. and Heo, T.-R., Survival of Bifidobacterium longum immobilized in calcium alginate beads in simulated gastric juices and bile salt solution. App. Environ. Microbiol., 66, 869- 873 (2000) https://doi.org/10.1128/AEM.66.2.869-873.2000
  13. Lian, W.-C., Hsiao, H.-C., and Chou, C.-C., Survival of bifidobacteria after spray-drying, Int. J. Food Microbiol., 74, 79-86 (2002) https://doi.org/10.1016/S0168-1605(01)00733-4
  14. Louis, K., Upton, M., and McLoughlin, A., Enhancing the viability of Lactobacillus plantum inoculum by immobilizing the cells in calcium alginate beads incorporating cryoprotectants, Appl. Environ. Microbiol., 56, 3112-3116 (1990)
  15. Picot, A. and Lacroix, C., Effect of micronization on viability and thermotolerance of probiotic freeze-dried cultures. Int. Dairy J., 13, 455-462 (2003) https://doi.org/10.1016/S0958-6946(03)00050-5
  16. Rao, A. V., Shiwnarain, N., and Maharaj, J., Survival of microencapsulated Bifidobacterium pseudolongum in simulated gastric and intestinal juices. Canadian Institute of Food Science and Technology, 22, 345-349 (1989) https://doi.org/10.1016/S0315-5463(89)70426-0
  17. Rasic, J. L. and Kurmann, J. A., Bifidobacteria and their role: microbiological, nutritional-physiological, medical and technological aspects and bibliography (Exs (experientia supplementum)), Birkhauser-Verlag-Basel, Basel, Switzerland, pp. 3-7 (1983)
  18. Shah, N. P. and Ravula, R. R., Microencapsulation of probiotic bacteria and their survival in frozen fermented dairy desserts, Aust. J. Dairy Technol., 55, 139-144 (2000)
  19. Wang, Y.-C., Yu, R.-C., and Chou, C.-C., Viability of lactic acid bacteria and bifidobacteria in fermented soymilk after drying, subsequent rehydration and storage, Int. J. Food Microbiol., 93, 209-217 (2004) https://doi.org/10.1016/j.ijfoodmicro.2003.12.001