Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Enzyme Property of a Cell-Wall Lytic Enzyme Produced by Bacillus sp. LM-8 against Lactobacillus plantarum.

Bacillus sp. LM-8이 생산하는 Lactobacillus plantarum 용균 효소의 정제 및 효소 특성

  • 마호우 (강원대학교 바이오 산업공학부) ;
  • 신원철 (강원대학교 바이오 산업공학부)
  • Published : 2002.03.01
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Abstract

Purification and characterization of enzyme property of a cell-wall lytic enzyme against Lactobacillus plantarum were carried out. Final specific activity of purified enzyme was 5.8 units/mg and purity of the enzyme was increased 8.3 fold compared with the enzyme activity in culture broth. The molecular weight of purified enzyme was estimated to be 60,000 kDa by gel filtration and SDS-polyacrylamide gel electrophoresis. Optimal pH and temperature for the activity of this enzyme were 3.0 and 4$0^{\circ}C$, respectively. The cell-wall lytic enzyme activity was maintained at 3$0^{\circ}C$ when treating the enzyme for 30 mins, whereas the activity was decreased to 80% of the maximum level at 4$0^{\circ}C$ The enzyme activity exhibited good stability at the range of pH 4~7.

Lactobacillus plantarum 용균 효소를 생산하는 균주를 배양하여 생산된 효소를 정제한 결과 비활성도가 5.8 units/mg protein 이었고 정제도 8.3배, 수율은 30%이었다. 정제효소의 분자량은 gel filtration과 SDS-polyacrylamide gel eletrophoresis를 이용하여 측정한 결과 60,000 kDa 이었다. 용균 효소의 최적 반응 시간은 20분이었으며 최적 온도는 4$0^{\circ}C$, 최적 pH는 3.0이었다. 온도 안정성은 각 온도에서 30분간 처리하였을 때 $30^{\circ}C$까지는 안정하였으나 4$0^{\circ}C$에서는 80% 활성을 나타내었다. 효소의 pH 안정성은 실온에서 1시간 처리하였을 때 pH 4~7에서 안정성을 나타내었다.

Keywords

References

  1. Buchanan, R. E. and N. E. Gibbson. 1974. Bergey's Manual of Determinative Bacteriology, Pp. 529-550. 8th ed. Williams and Wilkins, Baltimore.
  2. Cooper, T. G. 1977. The Tools of Biochemistry, p. 169-193. A Wiley-Interscience, Publication, New York.
  3. Funatsu, M. and D. Tsuru. 1977. Youkin kouso (lytic enzyme). Koudansha Co. Tokyo Japan.
  4. Hayashi, K., T. Kasumi, N. Kubo, and N. Tsumura. 1984. Properties of N-acetylmuramidase from Streptomyces rutgersensis H-46.Agric. Bioi. Chem. 48: 465-471.
  5. Hayashi, K., T. Kasumi, N. Kubo, K. Haraguchi, and N. Tsumura. 1989. Effects of N-acetylmuramidase from Streptomyces rutgersensis H-46 as a food preservative. Agric. BioI. Chem. 39: 2305-2316.
  6. Hideo, M., I. Morishima, and T. Ueno. 1991. Lytic enzyme for latic acid bacteria produced by Aspergillus cellulosae. J. Ferment. Technol. 69: 75-82.
  7. Hunghy, V L. and E. A. Johnson. 1987. Antimicrobial activity of lysozyme against bacteria involved in food spoilage and food-borne disease. Appl. Environ. Microbiol. 53: 2165-2170.
  8. Hunghy, V L., P. A. Wilger, and E. A. Johnson. 1989.Antimicrobial activity of hen egg white lysozyme against Listeria monocytogenes scott in foods. Appl. Environ. Microbiol. 55: 631-638.
  9. Janson, J. C. and L. Ryden. 1989. Protein Purification, Pp. 63-106. VCH Publishers, New York.
  10. Jung, M. H., I. S. Kong, D. H. Bai, and J. H. Yu. 1991. Purification and characterization of a bacteriolytic enzyme from alkalophilic Bacillussp. J Microbial. Biotech. 1: 102-110.
  11. Kim, Y. M., B. H. Lee, S. H. Lee, I. S. Shin, and T. S. Lee. 1988. The preservative effect of egg white lysozyme added surumiproducts. Bull. Korean. Fish. Soc. 21: 269-275.
  12. Laemmli, U. K. and M. Favre. 1973. Maturation of the head of bacteriophage T4. J Mol. Bioi. 80: 575-599.
  13. Lee, S. K., S. P. Hong, Y. B. Kim, S. Y. Choi, and I. J. Yoo. 1991. Effect of lysozyme and sodium ultraphosphate on the lysis of L. plantarum. Korean J Anim. Sci. 33: 73-77.
  14. Murao, S. and Y. Takahara. 1974. Enzyme lytic against Pseudomonas aeruginosa produced by Bacillus subtilis YT26. Agric. Bio!. Chem. 39: 2305-2316.
  15. Samuelson, K. J., J. H. Rupnow, and G. W. Froning. 1985. The effect of lysozyme and ethylenediaminetetraacetic acid on Samonella on broiler parts. Poultry Science. 64: 1488-1490.
  16. Scopes, R K. 1987. Protein purification, p. 186-198. SpringerVerlag, New York.
  17. Shin, W. C. and H. W. Ma. 1996.Isolationand identification of a lytic enzyme producing microorganism against lactic acid bacteria and its culture condition. Kor. J. Appl. Microbio!. Biotechnol. 24: 299-303.
  18. Strominger, J. L. and 1. M. Ghuysen. 1967. Mechanism of enzymatic bacteriolysis. Science. 156: 213-221.
  19. Uchida, M., S. Yokomura, and G. Nagahama. 1972. Studies on Lactobacilli isolationfrom mirin liquor. J. Ferment. Technol 50: 292-297.
  20. Walker, J. M. 1984. Protein, Pp. 41-55. Humana Press. Clifton, New Jersey.
  21. Yajima. M., Y. Hidaka, and Y. Matsuoka. 1968. Studies on egg white lysozyme as a preservative of sake. J. Ferment. Technol. 46: 782-788.
  22. Yokogawa, K., S. Kawata, and Y. Yoshimura. 1973. Lytic enzyme from Streptomyces globisporus. Agric. Bioi. Chem. 37: 799-808.