Optimization of Culture Conditions and Encapsulation of Lactobacillus fermentum YL-3 for Probiotics

가금류 생균제 개발을 위한 Lactobacillus fermentum YL-3의 배양조건 최적화 및 캡슐화

  • Kim, Kyong (Department of Food Science and Technology, Chungbuk National University) ;
  • Jang, Keum-Il (Department of Food Science and Technology and Research Center for Bioresource and Health, Chungbuk National University) ;
  • Kim, Chung-Ho (Department of Food and Nutrition, Seowon University) ;
  • Kim, Kwang-Yup (Department of Food Science and Technology and Research Center for Bioresource and Health, Chungbuk National University)
  • 김경 (충북대학교 식품공학과) ;
  • 장금일 (충북대학교 식품공학과 및 생물건강산업개발연구센터) ;
  • 김정호 (서원대학교 식품영양학과) ;
  • 김광엽 (충북대학교 식품공학과 및 생물건강산업개발연구센터)
  • Published : 2002.04.01

Abstract

This experiment was performed to improve the stability of Lactobacillus fermentum YL-3 as a poultry probiotics. The culture conditions that improve acid tolerance of L. fermentum YL-3 were investigated by changing several factors such as medium composition, temperature, anaerobic incubation and culture time. Also, L. fermentum YL-3 was encapsulated with alginate, calcium chloride and chitosan. The stable culture conditions of L. fermentum YL-3 were obtained in anaerobic incubation using MRS media without tween 80 for 20 hour at $42^{\circ}C$. The capsule after treatment with 1% chitosan was formed close membrane by a bridge bond. Immobilization of L. fermentum YL-3 in capsule was observed by confocal laser scanning microscopy, and cell viability was $2.0{\times}10^9\;CFU/g$ above the average. L. fermentum YL-3 capsule after acid treated at pH 2.0 for 3 hour survived about 40%, but those encapsulated with 1% chitosan survived about 65%. Survival rate of capsule stored at room temperature decreased about $2{\sim}3$ log cycle during 3 weeks, but viability of capsule stored at $4^{\circ}C$ during 3 weeks maintained almost $10^8\;CFU/g$ levels.

Keywords

probiotics;Lactobacillus fermentum YL-3;acid tolerance;encapsulation

References

  1. Conway, P.L. and Goldin, B.R. Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells. J. Dairy Sci.70: 1-12 (1987) https://doi.org/10.3168/jds.S0022-0302(87)79974-3
  2. Mcdonald, L.C., Fleming, H.P. and Hassan, H.M. Acid tolerance of Leuconostoc mesenteriodes and Lactobacillus plantarum. Appl.Environ. Microbiol. 56: 2120-2124 (1990)
  3. Nikkila, P, Johnsson, T., Rosenqvist, H. and Toivonen, L. Effect of pH on growth and fatty acid composition of Lactobacillus buchneri and Lactobacillus fermentum. Appl. Biochem. Biotech.59: 245-258 (1996) https://doi.org/10.1007/BF02783568
  4. Bartkowiak, A. Alginate-oligochitosan microcapsules. II. Control of mechanical resistance and permeability of the membrane, Chem. Mater. 12: 206-212 (2000) https://doi.org/10.1021/cm991128m
  5. Mundt, J.O. Enterococci; In Bergey's manual of systematic Bacte-riology, Vol. 2, p. 1063, Williams & Wilkins, London, (1986)
  6. Cho, Y.H., Shin, D.S. and Park, J.Y. Microencapsulation technol-ogy of food industry. Food Sci. Ind. 30: 98-111 (1997)
  7. MID. Inc. Operating manual Ver 6, Sherlock microbial identifica-tion system
  8. Shin, M.S., Kim, H.M., Kim, G.T., Huh, C.S., Bae, H.S. and Baek, Y.J. Selection and characteristic of Lactobacillus acidophilus isolated from Korean feces. Korean J. Food Sci. Technol. 31:495-501 (1999)
  9. Dziezak, J.D. Microencapsulation and Encapsulated Ingredient.Food Technol. 42: 136 (1988)
  10. Chang, H.N. and Seong, G.H. Microencapsultion of recombinant Saccharomyces cerevisiae cells with invertase activity in lipid-core alginate capsules. Biotech. Bioeng. 51: 157-162 (1996) https://doi.org/10.1002/(SICI)1097-0290(19960720)51:2<157::AID-BIT4>3.0.CO;2-I
  11. Promspone, B., Morishita, T.Y., Aye, P.P, Cobb, C.W., Reldkamp,A. and Clifford, J.R. Evaluation of avian specific probiotic and Salmonella typhimurium-specific antibodies on the colonization of Salmonella typhimurium in broiler. J. Food Protec. 61: 176-180(1998) https://doi.org/10.4315/0362-028X-61.2.176
  12. Cho, M.K., Kim, K., Kim, C.H. and Kim, K.Y. Isolation and characterization of Lactobacillus fermentum YL-3 as a poultry probiotic. Korean J. Appl. Microbiol. Biotechnol. 28: 279-284(2000)
  13. Sim, J.H., Kim, S.K., Baek, Y.J., Oh, T.K. and Yang, H.C. Influ-ence of culture conditions on acid tolerance of Lactobaciltus casei YIT 9018. Korean J. Appl. Microbiol. Biotechnol. 23: 17-23 (1995)
  14. Kleanhammer, T.R. Microbiological consideration in selection and preparation of Lactobacillus strains for use as dietary. J. Dairy Sci. 65: 1339-1349 (1982) https://doi.org/10.3168/jds.S0022-0302(82)82351-5
  15. Fuller, R. Probiotics in man and animal, J. Appl. Bacteriol. 66:365-378 (1989) https://doi.org/10.1111/j.1365-2672.1989.tb05105.x
  16. Park, C.J., Pyeon, J.S., Cho, Y.K., Hong, S.S. and Lee, H.S. Characteristics of Enterococcus sp. isolated from animal intestine and its powder. Korean J. Appl. Microbiol. Biotechnol. 24: 393-398(1996)
  17. Hassan, A.N. and Frank, T.F. Observation of encapsulated lactic caid bacteria using confocal laser microscopy, J. Dairy Sci. 78:2624-2628 (1995) https://doi.org/10.3168/jds.S0022-0302(95)76891-6
  18. Johnsson, T., Nikkila, P, Toivonen, L., Rosenqvist, H. and Laakso, S. Cellular fatty acid profiles of Lactobacillus and Lacto-coccus strains in relation to the oleic acid content of the cultiva-tion medium. Appl. Environ. Microbiol. 61: 4497-4499 (1995)
  19. Kim, H.S. Studies on the viability of Lactobacillus acidophitus IFO 3205 by microencapsulation, Ph.D. dissertation, Seoul National University, Seoul (1990)
  20. Kim, J.G., Lee, S.H., Lee, C.H., Lee, N.S., Son, Y.S. and Lim, S.K. Field treatment of cow manure originated from the clay mineral feeding and the change of nitrogen in soils. Korean J. Environ. Agri. 18: 366-371 (1999)