Probiotic Property of Lactobacillus pentosus Miny-148 Isolated from Human Feces

인체분변으로부터 분리한 유산균 Lactobacillus pentosus Miny-148의 생균제 특성 연구

  • 정민영 (한국생명공학연구원 생물자원센터) ;
  • 박용하 (영남대학교 응용미생물학과) ;
  • 김현수 (충남대학교 수의과대학) ;
  • 부하령 (한국생명공학연구원 생물자원센터) ;
  • 장영효 (한국생명공학연구원 생물자원센터)
  • Received : 2009.02.27
  • Accepted : 2009.03.27
  • Published : 2009.06.30

Abstract

Three hundred lactic acid bacteria isolated from human feces were studied their probiotic characters to develop potential probiotics. The properties were tested on the basis of guideline for probiotic selection protocol such as tolerance for acid or bile salt, thermal stability, antimicrobial, anticancer cell, and antiviral activity. Strain Miny-148 was selected as a potential probiotic bacterium which showed resistance to low pH, bile salts and thermal stability. On the basis of fatty acid profiles and 16S rDNA sequences analysis, the strain was identified as Lactobacillus pentosus (similarity 99.9%). The strain, L. pentosus Miny-148, showed broad antimicrobial spectrum against E. coli O157:H7, Shigella flexneri, Bacillus anthracis, Staphylococcus aureus, E. coli, Vibrio cholerae, V. vulnificus, Salmonella typhimurium, and Methicillin-resistant S. aureus (MRSA). Cell-free culture supernatant of the strain also inhibited against the growth of HT-29 colon cancer cell and transmissible gastroenterits virus.

우수한 생균제를 개발하기 위하여 안전성이 알려진 유산균을 대상으로 인체의 분변으로부터 300여 균주를 분리하고 내산성, 내담즙성, 내열성, 항균력, 항암 및 항바이러스 효과를 가지는 균주들을 선발하여 생균제 특성을 나타내는지를 알아보기 위해 본 실험을 수행하였다. 인체에서 분리한 여러 균주 중 Miny-148 균주는 낮은 pH 및 높은 담즙산에 대한 내성, 열처리에 대한 열안정성을 지녀 기초적인 probiotic 특성을 가진 균주로 선발되어, Lactobacillus pentosus (99.9% 상동성)로 동정되었다. 항균력 실험에서 Escherichia coli O157:H7을 비롯한 Shigella flexneri, Bacillus anthracis, Staphylococcus aureus, E. coli, Vibrio cholerae, V. vulnificus, Salmonella typhimurium, 그리고 및 methicillin-resistant S. aureus (MRSA) 균주 8종 등 총 16종의 병원성세균을 억제하였다. 또한 Miny-148은 결장암 세포인 HT-29 cell을 억제하였을 뿐 아니라, transmissible gastroenteritis virus의 생육을 저해하여 세포변성 억제효과를 가진 우수한 probiotic 특성을 지닌 균주로 분석되었다.

Keywords

References

  1. Baricault, L., G. Denariaz, J.J. Houri, C. Bouley, C. Sapin, and G. Trugnan. 1995. Use of HT-29, a cultured human colon cancer cell line, to study the effect of fermented milks on colon cancer cell growth and differentiation. Carcinogenesis 16, 245-252 https://doi.org/10.1093/carcin/16.2.245
  2. Berg, R.D. 1998. Probiotics, prebiotics or conbiotics. Trends Microbiol. 6, 89-92 https://doi.org/10.1016/S0966-842X(98)01224-4
  3. Bezkorovainy, A. 2001. Probiotics: determinants of survival and growth in the gut. Am. J. Clin. Nutr. 73, 399S-405S https://doi.org/10.1093/ajcn/73.2.399s
  4. Byun, J.W., G.T. Kim, H.S. Bae, Y.J. Baek, and Y.K. Lee. 2000. In vitro selection of lactic acid bacteria for probiotic use in pigs. Korea J. Vet. Res. 40, 701-706
  5. Casey, P.G., G.D. Casey, G.E. Gardiner, M. Tangney, C. Stanton, R.P. Ross, C. Hill, and G.F. Fitzgerald. 2004. Isolation and characterization of anti-Salmonella lactic acid bacteria from the porcine gastrointestinal tract. Lett. Appl. Microbiol. 39, 431-438 https://doi.org/10.1111/j.1472-765X.2004.01603.x
  6. Chang, Y.H., J.K. Kim, H.J. Kim, W.Y. Kim, Y.B. Kim, and Y.H. Park. 2001. Selection of a potential probiotic Lactobacillus strain and subsequent in vivo studies. Antonie Van Leeuwenhoek 80, 193-199 https://doi.org/10.1023/A:1012213728917
  7. Chang, Y.H., J.K. Kim, H.J. Kim, J.H. Yoon, W.Y. Kim, Y.W. Choi, W.J. Lee, Y.B. Kim, and Y.H. Park. 1999. Characteristics of Lactobacillus reuterii BSA -131 isolated from swine intestine. Kor. J. Appl. Microbiol. Biotechnol. 27, 23-27
  8. Cho, M.K., K. Kim, C.H. Kim, T.K. Lee, and K.Y. Kim. 2000. Isolation and characterization of Lactobacillus fermentum YL-3 as a poultry probiotic. Kor. J. Appl. Microbiol. Biotechnol. 28, 279-284
  9. Foligne, B., S. Nutten, C. Grangette, V. Dennin, D. Goudercourt, S. Poiret, J. Dewulf, D. Brassart, A. Mercenier, and B. Pot. 2007. Correlation between in vitro and in vivo immunomodulatory properties of lactic acid bacteria. World J. Gastroenterol. 13, 236-243 https://doi.org/10.3748/wjg.v13.i2.236
  10. Gallagher, T.M. and M.J. Buchmeier. 2001. Coronavirus spike proteins in viral entry and pathogenesis. Virology 279, 371-374 https://doi.org/10.1006/viro.2000.0757
  11. Gram, L., J. Melchiorsen, B. Spanggaard, I. Huber, and T.F. Nielsen. 1999. Inhibition of Vibrio anguillarum by Pseudomonas fluorescens AH2, a possible probiotic treatment of fish. Appl. Environ. Microbiol. 65, 969-973
  12. Ha, C.G., J.K. Cho, Y.K. Chai, and K.C. Heo. 2004. Isolation and identification of lactic bacteria containing superior activity of the bile salts deconjugation. Kor. J. Food Sci. Ani. Resour. 24, 164-170
  13. Kim, K., K.L. Jang, C.H. Kim, and K.Y. Kim. 2002. Optimization of culture conditions and encapsulation of Lactobacillus fermentum YL-3 for probiotics. Kor. J. Food Sci. Technol. 34, 255-262
  14. Kim, P.I., M.Y. Jung, Y.H. Chang, S. Kim, S.J. Kim, and Y.H. Park. 2007. Probiotic properties of Lactobacillus and Bifidobacterium strains isolated from porcine gastrointestinal tract. Appl. Microbiol. Biotechnol. 74, 1103-1111 https://doi.org/10.1007/s00253-006-0741-7
  15. Kim, Y.G., J.T. Moon, K.M. Lee, N.R. Chon, and H. Park. 2006. The effects of probiotics on symptoms of irritable bowel syndrome. Korean J. Gastroenterol. 47, 413-419
  16. Koga, T., T. Mizobe, and K. Takumi. 1998. Antibacterial activity of Lactobacillus species against Vibrio species. Microbiol. Res. 153, 271-275 https://doi.org/10.1016/S0944-5013(98)80011-6
  17. Lee, E.Y. 2008. Problems and verification system of probiotics as livestock-environment improving agent produced and circulated. Kor. J. Microbiol. Biotechnol. 36, 87-95
  18. O'Sullivan, M.A. and C.A. O'Morain. 2000. Bacterial supplementation in the irritable bowel syndrome. A randomised double-blind placebo-controlled crossover study. Dig. Liver Dis. 32, 294-301 https://doi.org/10.1016/S1590-8658(00)80021-3
  19. Reed, M.J. and M. Muench. 1938. A simple method for estimating fifty percent endpoints. Am. J. Hygiene 27, 493-497
  20. Ringel-Kulka, T. and Y. Ringel. 2007. Probiotics in irritable bowel syndrome: has the time arrived? Gastroenterology 132, 813-816 https://doi.org/10.1053/j.gastro.2006.12.058
  21. Ringel, Y., A.D. Sperber, and D.A. Drossman. 2001. Irritable bowel syndrome. Annu. Rev. Med. 52, 319-338 https://doi.org/10.1146/annurev.med.52.1.319
  22. Rosenfeldt, V., E. Benfeldt, S.D. Nielsen, K.F. Michaelsen, D.L. Jeppesen, N.H. Valerius, and A. Paerregaard. 2003. Effect of probiotic Lactobacillus strains in children with atopic dermatitis. J. Allergy Clin. Immunol. 111, 389-395 https://doi.org/10.1067/mai.2003.389
  23. Saggioro, A. 2004. Probiotics in the treatment of irritable bowel syndrome. J. Clin. Gastroenterol. 38(6 Suppl), S104-106 https://doi.org/10.1097/01.mcg.0000129271.98814.e2
  24. Sartor, R.B. 2001. Intestinal microflora in human and experimental inflammatory bowel disease. Curr. Opin. Gastroenterol. 17, 324-330 https://doi.org/10.1097/00001574-200107000-00005
  25. Sartor, R.B. 2008. Microbial influences in inflammatory bowel diseases. Gastroenterology 134, 577-594 https://doi.org/10.1053/j.gastro.2007.11.059
  26. Shah, G.D., J.S. Silver, S.S. Rosenfeld, I.T. Gavrilovic, L.E. Abrey, and A.B. Lassman. 2007. Myelosuppression in patients benefiting from imatinib with hydroxyurea for recurrent malignant gliomas. J. Neurooncol. 85, 217-222 https://doi.org/10.1007/s11060-007-9408-1
  27. Shin, M.S., H.M. Kim, G.T. Kim, C.S. Huh, H.S. Bae, and Y.J. Baek. 1999. Selection and characteristics of Lactobacillus acidophilus isolated from Korean feces. Kor. J. Food Sci. Technol. 31, 495-501
  28. Sinn, D.H., J.H. Song, H.J. Kim, J.H. Lee, H.J. Son, D.K. Chang, Y.H. Kim, J.J. Kim, J.C. Rhee, and P.L. Rhee. 2008. Therapeutic effect of Lactobacillus acidophilus-SDC 2012, 2013 in patients with irritable bowel syndrome. Dig. Dis. Sci. 53, 2714-2718 https://doi.org/10.1007/s10620-007-0196-4
  29. Weese, J.S., M.E. Anderson, A. Lowe, R. Penno, T.M. da Costa, L. Button, and K.C. Goth. 2004. Screening of the equine intestinal microflora for potential probiotic organisms. Equine Vet. J. 36, 351-355 https://doi.org/10.2746/0425164044890616