Probiotic Characteristics of Lactobacillus rhamnosus Isolated from Kefir

Kefir로부터 분리한 Lactobacillus rhamnosus의 Probiotic 특성

  • 유숙진 (국립한경대학교 낙농과학과) ;
  • 조진국 (고품질친환경농축산물 생산기술연구센터) ;
  • 황성구 (국립한경대학교 낙농과학과) ;
  • 허강칠 (국립한경대학교 낙농과학과)
  • Published : 2005.09.01

Abstract

To search probiotic microorganisms, we isolated Lactobacillus sp. from kefir, The Lactobacillus sp. strain showed $99.5\%$ of identity to species Lactobacillus rhamnosus by API kit. Lactobacillus rhamnosus showed high resistances to acidic environment, which grew well even at pH 2.0 and $1.0\%$ bile salt Enzyme activity of Lactobacillus rhamnosus was higher in amylase ($0.673\;{\mu}mol/min/mg$) than that in xylanase ($0.288\;{\mu}mol/min/mg$), cellulase($0.117\;{\mu}mol/min/mg$) and phytase($0.269\;{\mu}mol/min/mg$). Especially, the Lactobacillus rhamnosus showed high heat stability which remained $1{\times}10^6\;CFU/ml$ at $60^{\circ}C$. The maximum numbers of Lactobacillus rhamnosus on growth owe was reached at 24 h fermentation and pH was decreased to 4.6. The resistances of Lactobacillus rhamnosus to acidic pH and bile salt were better than that of Lactobacillus acidophilus used as control. When Lactobacillus rhamnosus was cultured with E. coli in MRS broth, E. coli was disappeared after 18 h. These result suggest that the isolated Lactobacillus rhamnosus has a useful probiotics properties.

References

  1. APHA (1985) Standard methods for the examination of dairy products, 15th ed, American Public Health Association, Washington. D. C
  2. Booth, I. R. (1985) Regulation of cytoplasmic pH in bacteria. Microbiol. Rev. 49, 359-378
  3. Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  4. Fuller, R. and Gibson, G. R. (1977) Modification of the intestinal microflora using probiotics and prebiotics. Scand. J. Gastroenterol. 222, 28-31
  5. Gilliand, S. E. and Speck, M. L. (1997) Deconjugation of bile acids by intestinal lactobacilli. Appl. Environ. Microbiol. 33, 15-18
  6. Gilliland, S. E. (1979) Benifical interrelationships between certain microorganisms and humans: candidate micro-organisms for use as dietary adjuncts. J. Food Prot. 42, 164-167 https://doi.org/10.4315/0362-028X-42.2.164
  7. Haenlein, G. F. W. (1995) Status and prospects of the dairy goat industry in the United States. J. Anim. Sci. 74, 1173-1181
  8. Havenaar, R., Brink, B. T., and Veld, J. H. (1992) Selection of strains for probiotic use. In 'Probiotics' Fuller, R.(ed), Chapman & Hall, New York, pp. 209-224
  9. Hood, S. K. and Zottola, E. A. (1988) Effect of low pH on the ability of Lactobacillus acidophilus to survive and adhere to human intestinal cells. J. Food Sci. 53, 1541-1516
  10. Itoh, K. (1990) Lactic acid bacteria and intestinal micro-flora, The 11th Internation Symposium on Lactic Acid Bacteria and Human Health, Seoul, Korea, pp. 23-25
  11. Khasin, A., Alchanati, I., and Shoham, Y. (1993) Purification and characterization of a thermostable xylanase from Bacillus stearothermophilus T-6. Appl. Environ. Microbiol. 59, 1725-1730
  12. Kimura, K., McCartney, A. L., McConell, M. A., and Tannock, G. W. (1977) Analysis of fecal populations of bifidobacteria and lactobcilli and investigation of the immunological response of their human hosts to the predominant strains. Appl. Environ. Microbiol. 63, 3394-3398
  13. Klaver, F. A. M. and van der Meer, R. (1993). The assumed assimilation of cholesterol by Lactobacillus and Bifidobacterium bifidum is due to their bile salt deconjugation activity. Appl. Environ. Microbial. 59, 1120-1124
  14. Koroleva, N. S. (1988). Technology of kefir and kumys. Internation Dairy Federation Bulletin. 227, 96-99
  15. Kobayashi, Y., Tohyaman, K., and Terashima, T. (1974) Studies on biological characteristics of Lactobacillus. II. Tolererance of the multiple antibiotic resistance-strain, L. casei PSR3002, to artificial digestive fluids. Japan. J. Microbio. 29, 691-697
  16. Kroger, M. and Kurmann, J. A. (1989) Fermented milks - past. present and future. Food Technol. 43, 92-99.
  17. Loffler, F. E., Sun, Q., Li, J., and Tiedje, J. (2000) 16S rRNA gene-base detection of tetrachoroethene-dechlorinating desulfuromonase and dehaloccoides species. Appl. Enviroron. Microbiol. 66, 1369-1374 https://doi.org/10.1128/AEM.66.4.1369-1374.2000
  18. Otle', S. and Cagindi, O. (2003) Kefir; A probiotic dairy-composition, nutritional and therapeutic aspects. Pakistan J. of Nutrition 2(2), 54-59 https://doi.org/10.3923/pjn.2003.54.59
  19. Park, S. Y., Ko, Y. T., Jeong, H. K., Yang, J. O., Chung, H. S., Kim, Y. B., and Ji, G. E. (1996) Effect of various lactic acid bacteria on the serum cholesterol levels in rats and resistance to acid bile and antibiotics. Kor. J. Appl. Microbiol. Biotechnol. 24, 304-310
  20. Shiomi, M. K., Sakai, M., Murofushi, M., and Aibara, K. (1982) Antitumor activity in mice of orally administered polysaccharide from Kefir grain. Jpn. J. Med. Sci. Biol. 35(2), 75-80 https://doi.org/10.7883/yoken1952.35.75
  21. Shimizu, M. (1992) Purification and characterization of phytase from Bacillus subtilis(natto) N-77. Biosci. Biotech. Biochem. 56, 1266 https://doi.org/10.1271/bbb.56.1266
  22. Shiral, K., Revah-Molseev, S., Garcia-Garlbay, M., and Marshall, V. M. (1994) Ability of some strains of lactic acid bacteria bacteria to degrade phytic acid. Lett. Appl. Microbiol. 19, 366-369 https://doi.org/10.1111/j.1472-765X.1994.tb00477.x
  23. Smith, H. W. (1975) Persistence of tetracycline resistance in pig. Nature 258, 628-629 https://doi.org/10.1038/258628a0
  24. Tannock, G. W., Crichton, C., Welling, G. W., Koopman, J. P., and Midtvedt, T. (1988) Reconstitution of the gastrointestinal microflora of lactobacillus-free mice. Appl. Environ. Microbial. 54, 2971-2975
  25. Tannock, G. W. (1997) Probiotic properties of lactic acid bacteri: plenty of scope for fundamental R & D. Trends in Biotechnology. 15, 270-274 https://doi.org/10.1016/S0167-7799(97)01056-1
  26. Toba, T. (1987) Symposium Reports on Advance of Dairy Science and Technology in Japan. Japanese Journal of Dairy and Food Science 36(6), A235-A243
  27. Van Houweling, C. D. (1971) FDA's views regarding antibiotics used in feed. In: Proceedings of the university of Maryland nutrition conference, Washington, DC, MD, USA
  28. Walsh, G. A., Power, R. F., and Headon, D. R. (1993) Enzymes in the animal-feed industry. Tibtech. 11, 424-429 https://doi.org/10.1016/0167-7799(93)90006-U