Screening and Characterization of Lactate Dehydrogenase-producing Microorganism

  • Sung, Ha Guyn (Dept. of Food Biotechnology, Sungkyunkwan University) ;
  • Lee, Jae Heung (Institute of Life Science and Technology, Sungkyunkwan University) ;
  • Shin, Hyung Tai (Dept. of Food Biotechnology, Sungkyunkwan University)
  • Received : 2003.12.31
  • Accepted : 2004.05.18
  • Published : 2004.10.01


The objective of this work was to isolate a microorganism, able to produce high lactate dehydrogenase (LDH) activity, for use as a microbial feed additive. The LDH is an important enzyme for lactate conversion in the rumen, thereby possibly overcoming lactic acidosis owing to sudden increases of cereal in the diets of ruminants. In the present study, various bacterial strains were screened from a variety of environments. Among the isolated microorganisms, strain FFy 111-1 isolated from a Korean traditional fermented vegetable food called Kimchi showed the highest enzyme activity, along with retaining strong enzyme activity even in rumen fluid in vitro. Based on morphological and biochemical characteristics as well as compositions of cellular fatty acids plus API analyses, this strain was identified as Lactobacillus sp. The optimum temperature and pH for growth were found to be 30$^{\circ}C$ and pH 6.5, respectively. A maximum cell growth of 2.2 at $A_{650}$ together with LDH activity of 2.08 U per mL was achieved after 24 h of incubation. Initial characterization of FFy 111-1 suggested that it could be a potential candidate for use as a direct-fed microbial in the ruminant animals.


Supported by : Ministry of Agriculture and Forestry


  1. Aslan, V., S. M. Thamsborg, R. J. Jorgensen and A. Basse. 1995. Induced acute ruminal acidosis in goats treated with yeast (Saccharomyces cerevisiae) and biocarbonate. Acta Vet. Scand. 36:65-77.
  2. Bae, J. S., J. R. Byun and Y. H. Yoon. 2003. In vivo antagonistic effect of Lactobacillus helveticus CU 631 against Salmonella enteritidis KU 101 infection. Asian-Aust. J. Anim. Sci. 16:430-434.
  3. Baldwin, R. L. and L. P. Milligan. 1964. The purification of a pyridine nucleotide independent lactate dehydrogenase from the anaerobic bacterium. Biochim. Biophys. Acta, 92:421-428.
  4. Bergmeyer, H. U. 1974. Methods of Enzymatic Analysis, 3rd edn. Verlag Chemie Weinheim Academic Press, Inc., New York.
  5. Bradford, M. M. 1976. A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248-254.
  6. Chaucheyras, F., G. Fonty, G. Bertin, J. M. Salmon and P. Gouet. 1996. Effects of a strain of Saccharomyces cerevisiae (Levucell SC1), a microbial additive for ruminant, on lactate metabolism in vitro. Can. J. Microbiol. 42:927-933.
  7. Collins, C. H. and P. M. Lyne. 1976. Microbiological Methods. Butterworths, London, pp.169-180.
  8. Fleet, G. H., S. Lafon-Lafourcade and P. Ribereau-Gayon. 1984. Evaluation of yeast and lactic acid bacteria during fermentation and storage of Bordeaux wines. Appl. Environ. Microbiol. 48:1034-1038.
  9. Garfin, D. E. 1990. One-dimensional gel electrophoresis. In: Guide to protein purification (Ed. M. P. Deutscher), Academic Press, San Diego, USA. pp. 425-440.
  10. Garvie, E. I. 1980. Bacterial lactate dehydrogenases. Microbiol. Rev. 44:106-139.
  11. Gerhardt, P., R. G. E. Murray, R. N. Costilow, E. W. Nester, W. A. Wood, N. R. Krieg and G. B. Phillips. 1981. Manual of Methods for General Bacteriology, American Society for Microbiology, Washington, USA, pp. 410-441.
  12. Gilmour, M., H. J. Flint and W. J. Mitchell. 1994. Multiple lactate dehydrogenase activities of the rumen bacterium Selenomonas ruminantium. Microbiology, 140:2077-2084.
  13. Hino, T. and S. Kuroda. 1993. Presence of lactate dehydrogenase and lactate racemase in Megasphera elsdenii grown on glucose or lactate. Appl. Environ. Microbiol. 59:255-259.
  14. Hirano, M., H. Mori, Y. Miura, N. Matsunaga, N. Nakamura and T. Matsunaga. 1990. $\gamma$-Linolenic acid production by microalgae. Appl. Biochem. Biotechnol. 24/25:183-191.
  15. Hong, S. I., Y. J. Kim and Y. R. Pyun. 1999. Acid tolerance of Lactobacillus plantarum, Lebensm.-Wiss. u.-Technol. 32:142-148.
  16. Kim, D. H., H. P. Song, H. S. Yook, Y. G. Ryu and M. W. Byun. 2004. Isolation of enteric pathogens in the fermentation process of Kimchi (Korean fermented vegetables) and its radicidation by gamma irradiation. Food Control, 15:441-445.
  17. Kim, D. H., H. P. Song, H. S. Yook, Y. G. Ryu and M. W. Byun. 2004. Isolation of enteric pathogens in the fermentation process of Kimchi (Korean fermented vegetables) and its radicidation by gamma irradiation. Food Control, 15:441-445.
  18. Kung, L. Jr. and A. O. Hession. 1995. Prevention in vitro lactate accumulation in ruminal fermentation with Megasphaera elsdenii. J. Anim. Sci. 73:250-256.
  19. Laemmli, U. K. 1970. Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature (London), 227:680-785.
  20. J. H., Y. B. Lim, K. M. Park, S. W. Lee, S. Y. Baig and H. T. Shin. 2003. Factors affecting oxygen uptake by yeast Issatchenkia orientalis as microbial feed additive for ruminants. Asian-Aust. J. Anim. Sci. 16:1011-1014.
  21. Martin, S. A. and D. J. Nisbet. 1992. Effect of direct-fed microbials on rumen microbial fermentation. J. Dairy Sci. 75:1736-1744.
  22. Moore, L. V. H., D. M. Bourne and W. E. C. Moore. 1994. Comparative distribution and taxonomic value of cellular fatty acids in thirty three genera of anaerobic Gram-negative bacilli. Int. J. Syst. Bacteriol. 44:338-347.
  23. Peter, H. A. S., S. M. Nicholas, M. E. Harpe and J. G. Holt. 1986. Bergey's Manual of Systemetic Bacteriology, Vol. 2, Williams and Willkins Co., USA. pp. 1104-1139.
  24. Russell, J. B. and T. Hino. 1985. Regulation of lactate production in Streptococcus bovis: A spiraling effect that contributes to rumen acidosis. J. Dairy Sci. 68:1712-1721.
  25. Russell, J. B. and J. L. Rychlik. 2001. Factors that alter rumen microbial ecology. Science 292:1119-1122.
  26. Scheifinger, C. C., M. J. Latham and M. J. Wolin. 1975. Relationship of lactate dehydrogenase specificity and growth rate to lactate metabolism by Selenomonas ruminantium. Appl. Microbiol. 30:916-921.
  27. Slyter, L. L. 1976. Influence of acidosis on rumen function. J. Anim. Sci. 43:910-929.
  28. Yun, J. W. and D. H. Kim. 1998. A comparative study of mannitol production by two lactic acid bacteria, J. Ferment. Bioeng. 85:203-208.