Production of Trans-10, Cis-12 Conjugated Linoleic Acid by Megasphaera Elsdenii YJ-4: Physiological Roles in the Rumen

  • Kim, T.W. (Department of Animal Science, Cornell University) ;
  • Choi, N.J. (National Livestock Research Institute, RDA) ;
  • Hwangbo, J. (National Livestock Research Institute, RDA) ;
  • Hsu, Jih-Tay (Department of Microbiology, Cornell University) ;
  • Lee, Sang S. (Genebiotech Co., Ltd.) ;
  • Song, M.K. (Department of Animal Science, Chungbuk National University) ;
  • Seo, I.J. (Wootech, Inc.) ;
  • Kim, Y.J. (Department of Food and Biotechnology, Korea University)
  • Received : 2005.01.11
  • Accepted : 2005.04.22
  • Published : 2005.10.01


Megaspahera elsdenii YJ-4, which was previously isolated as a producer of trans-10, cis-12 CLA, was studied for its carbon source on the CLA production. M. elsdenii YJ-4, was incubated with glucose and lactose, and cultured in batch and continuous culture systems with linoleic acid at various pHs to investigate CLA production. Batch cultures of the ruminal bacterium, M. elsdenii YJ-4, were resistant to stearic acid and linoleic acid, and little growth inhibition was observed even when the fatty acid concentration in the culture was as much as 4 mg $ml^{-1}$. Stationary phase batch cultures (0.25 mg bacterial protein $ml^{-1}$) that had been grown on lactate and incubated with linoleic acid (0.20 mg $ml^{-1}$) produced approximately 12 ${\mu}g$ trans-10, cis-12 CLA mg $protein^{-1}$ and little cis-9, trans-11 CLA was detected. Some linoleic acid was converted to hydrogenated products (chiefly stearic acid), but these fatty acids were less than 5 ${\mu}g$ mg bacterial $protein^{-1}$. Stationary phase batch cultures that had been grown on glucose produced at least 3-fold less trans-10, cis-12 CLA than ones grown on lactate. Cells from lactate-limited continuous cultures produced less trans-10, cis-12 CLA than those from batch culture, but only if the pH was greater than 6.4. When the pH of the lactate-limited continuous cultures was lower than 6.4, trans-10, cis-12 CLA and hydrogenated products declined. Cells from glucose-limited continuous cultures produced less trans-10, cis-12 CLA and hydrogenated products than the cells that had been limited by lactate, but pH had little impact on this production. These results support the idea that M. elsdenii YJ-4 could be one of the major producers of trans-10, cis-12 CLA which causes cows to produce milk with a low fat content.


Biohydrogenation;Conjugated Linoleic Acid;Fatty acid;Megasphaera elsdenii;Rumen


Supported by : Rural Development Association


  1. Choi, B., S. Kang, Y. Ha, G. Park and R. G. Ackman. 2002. Conjugated linoleic acid as a supplemental nutrient for common carp (Cyprinus carpio). Food Sci. Biotechnol. 11:457-461.
  2. Dougan, M. E. R., J. L. Aalhus, A. L. Schaefer and J. K. G. Kramer. 1997. The effect of conjugated linoleic acid on fat to lean repartitioning and feed conversion in pigs. Can. J. Anim. Sci. 77:723-725.
  3. Henderson, C. 1973. The effects of fatty acids on pure cultures of rumen bacteria. J. Agric. Sci. 81:107-112.
  4. Hungate, R. E. 1966. The rumen and its microbes. Academic Press, New York.
  5. Kim, Y. J. and R. H. Liu. 1999. Selective increase in conjugated linoleic acid in milk fat by crystallization. J. Food Sci. 64:792-795.
  6. Kim, Y. J., R. H. Liu, D. Bond and J. B. Russell. 2000. The effect of linoleic acid concentration on the conjugated linoleic acid (CLA) production of Butyrivibrio fibrisolvens A38. Appl. Environ. Microbiol. 66:5226-5230.
  7. Kim, Y. J. 2003. Partial inhibition of biohydrogenation of linoleic acid can increase the conjugated linoleic acid production of Butyrivibrio fibrisolvens A38. J. Agric. Food Chem. 51:4258-4262.
  8. Stackebrandt, E., H. Pohla, R. Kroppenstedt, H. Hippe and C. R. Woese. 1985. 16S rRNA analysis of Sporomusa, Selenomonas, and Megasphaera: on the phylogenetic origin of Grampositive eubacteria. Arch. Microbiol. 143:270-276.
  9. Wang, J. H., S. H. Choi and M. K. Song. 2003. Effects of concentrate to roughage ratio on the formation of trans-11-$C_{18:1}$ and cis-9, trans-11-$C_{18:2}$ in rumen fluid and plasma of sheep when fed high oleic or high linoleic acid oil. Asian-Aust. J. Anim. Sci. 16:1604-1609.
  10. Callaway, T. R., K. A. Adams and J. B. Russell. 1999. The ability of 'low G+C gram-positive' ruminal bacteria to resist monensin and counteract potassium depletion. Curr. Microbiol. 39:225-230.
  11. Kim, Y. J., R. H. Liu, J. L. Rychlik and J. B. Russell. 2002. The enrichment of a ruminal bacterium (Megasphaera elsdenii YJ-4) that produces the trans-10, cis-12 isomer of conjugated linoleic acid (CLA) J. Appl. Microbiol. 92:976-982.
  12. Lee, K. N., D. Kritchevsky and M. W. Pariza. 1994b. Conjugated linoleic acid and atherosclerosis in rabbits. Atherosclerosis. 108:19-25.
  13. Wolin, M. J., T. L. Miller and C. S. Stewart. 1997. The Rumen Microbial Ecosystem (Ed. P. N. Hobson and C. S. Stewart). Blackie Academic & Professional, London, UK. pp. 467-491.
  14. Davis, C. L., R. E. Brown and D. C. Beitz. 1964. Effect of feeding high-grain restricted-roughage rations with and without bicarbonates on the fat content of milk produced and proportions of volatile fatty acids. J. Dairy Sci. 47:1217-1226.
  15. Ha, Y. L., N. K. Grimm and M. W. Pariza. 1987. Anticarcinogens from fried ground beef: heat-altered derivatives of linoleic acid. Carcinogenesis 8:1881-1888.
  16. Wang, J. H. and M. K. Song. 2003. pH affects the in vitro formation of cis-9, trans-11 CLA and trans-11 octadecenoic acid by ruminal bacteria when incubated with oilseeds. Asian-Aust. J Anim. Sci. 16:1743–1748.
  17. Kepler, C. R. and S. B. Tove. 1967. Biohydrogenation of unsaturated fatty acids. J. Biol. Chem. 242:5686-5692.
  18. Thompson, L., A. Cockayne and R. C. Spiller. 1994. Inhibitory effect of polyunsaturated fatty acids on the growth of Helicobacter pylori: a possible explanation of the effect of diet on peptic ulceration. Gut. 35:1557-1561.
  19. Baldwin, R. L., W. A. Wood and R. S. Emery. 1965. Lactate metabolism by Peptostreptococcus elsdenii: Evidence for lactyl coenzyme A dehydrase. Biochim. Biophy. Acta. 97:202-213.
  20. Brockman, H. L. and W. A. Wood. 1975. Acrylyl-coenzyme A. J. Bacteriol. 123:1447-1453.
  21. Dewhurst, R. J., D. Wadhwa, L. P. Borgida and W. J. Fisher. 2001. Rumen acid production from dairy feeds. 1. Effects on Feed intake and milk production of dairy cows offered grass or corn silages. J. Dairy Sci. 84:2721-2729.
  22. Lee, H. S., E. J. Kim, J. S. Cho, H. J. Cho, H. J. Cho, S. K. Lee, K. C. Jung and J. H. Y. Park. 1994a. Conjugated Linoleic Acid Decreases Phosphorylated Akt Levels in Mat-LyLu (MLL) Rat Prostate Cancer Cells. Food Sci. Biotechnol. 13:353-357.
  23. Gaynor, P. J., R. A. Erdman, B. B. Teter, J. Sampugna, J., A. V. Capuco, D. R. Waldo and M. Hamsosh. 1994. Milk fat yields and composition during abomasal infusion of Cis or Trans octadecenoates in Holstein cows. J. Dairy Sci. 77:157-165.
  24. Miwa, T., H. Esaki, J. Umemori and T. Hino. 1997. Activity of H(+)-ATPase in ruminal bacteria with special reference to acid tolerance. Appl. Environ. Microbiol. 63:2155-2158.
  25. Baumgard, L. H., J. K. Sangster and D. E. Bauman. 2001. Milk fat synthesis in dairy cows is progressively reduced by increasing supplemental amounts of trans-10, cis-12 conjugated linoleic acid (CLA). J. Nutr. 131:1764-1769.
  26. Counotte, G. H. M., R. A. Prins, R. H. A. M. Janssen and M. J. A. deBie. 1981. The role of Megasphaera elsdenii in the fermentation of DL-(2-13C)-lactate in the rumen of dairy cattle. Appl. Environ. Microbiol. 42:649-655.

Cited by

  1. Shifts in bacterial community composition in the rumen of lactating dairy cows under milk fat-depressing conditions vol.93, pp.1, 2010,