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전통 장류 유래 Bacillus subtilis SCJ4의 특성확인 및 통계학적 방법을 이용한 배양조건 최적화

Identification of characterization and statistical optimization of medium constituent for Bacillus subtilis SCJ4 isolated from Korean traditional fermented food

  • 투고 : 2015.02.05
  • 심사 : 2015.02.26
  • 발행 : 2015.03.31

초록

순창군에서 제조된 전통 장류로부터 612종의 다양한 미생물을 분리하였고, 612종의 미생물 중에서 생리활성 분석 및 biogenic amine 생성여부를 조사하여 최종적으로 Bacillus subtilis SCJ4를 선별하였다. 최종 선별한 균주 SCJ4의 생화학적 특성과 16S rRNA 염기서열 분석에 의해 B. subtilis SCJ4로 명명하였으며, B. subtilis SCJ4의 산업적으로 사용이 적합한 최적배지조성을 확립하기 위하여 통계학적 방법인 반응표면 분석법을 사용하였다. 실험계획법은 배지조성을 예측하기 위해 Plackett-Burman design을 사용하여 모델을 설정하였다. 실험 모델을 통하여 선별된 배지 조성으로는 tryptone, peptone, $MgSO_4$로 예측되었으며, 배지조성에 대한 농도를 설정하기 위해 중심합성계획법을 사용하여 최적화를 수행하였다. 최종 예측된 배지 조성의 농도는 tryptone 15.35 g/L, peptone 12.235 g/L, $MgSO_4$ 3.5 g/L로 예측되었다. 이때 최대 균체량은 1.07497 g/L로 예측되었으며, 모델의 검증실험을 통하여 예측된 모델과 실제 결과가 동일함을 확인하였으며, 기본배양배지와 비교하였을 때 균체량이 0.8767 g/L에서 1.1222 g/L으로 약 28%증가하였다.

612 strains isolated from Korean traditional fermented food in Sunchang and their investigated biochemical characterization and ability of biogenic amines non-producing. We selected the SCJ4 having various activity by measurement of extracellular enzyme, antioxidant and antimicrobial activities. Selected strain SCJ4 by 16S rRNA sequencing and biochemical characterization was named Bacillus subtilis SCJ4. And then, we investigated cell growth of SCJ4, and optimized of culture medium constituents using response surface methodology as statistically method. Response surface methodology used Plackett-Burman experimental design for screening of medium constituent. Tryptone, peptone and $MgSO_4$ as medium constituent improving cell growth selected. In order to find out optimal concentration on each constituent, we carried out central composite design. Consequently, optimized concentrations of tryptone, peptone and $MgSO_4$ were predicted to be 15.35 g/L, 12.235 g/L, and 3.5 g/L respectively. Through the model verification, we confirmed about 1.28-fold improvement of the dried cell weight from 0.8767 g/L to 1.1222 g/L when compared to basal medium.

키워드

참고문헌

  1. Baek, S.Y., Lee, Y.J., Yun, H.J., Park, H.Y., and Yeo, S.H. 2014. Characterization of alkaline cellulase from Bacillu subtilis 4-1 Isolated from Korean traditional soybean paste. Korean J. Food Preserv. 21, 442-450. https://doi.org/10.11002/kjfp.2014.21.3.442
  2. Baek, S.Y., Yun, H.J., Choi, H.S., Koo, B.S., and Yeo, S.H. 2010. Isolation and physiological characteristics of microorganisms producing extracellular enzymes from Korean traditional soybean sauce and soybean paste. Korean J. Microbiol. Biotechnol. 38, 379-384.
  3. Cha, J.Y., Kim, H.J., Jun, B.S., Park, J.C., Ok, M., and Cho, Y.S. 2003. Antioxidative and produced condition of antioxidative substance by Bacillus sp. FF-7. J. Korean Soc. Agric. Chem. Biotechnol. 46, 165-170.
  4. Chang, M. and Chang, H.C. 2012. Development of a screening method for biogenic amine producing Bacillus spp. Int. J. Food Microbiol. 150, 269-274.
  5. Chi, W.J., Park, D.Y., Uyangaa, T., Lee, J.Y., Chang, Y.K., and Hong, S.K. 2011. Identification of a cellulase producing marine Bacillus sp., GC-1 and GC-4 isolated from coastal seawater of Jeju island. Korean J. Microbiol. Biotechnol. 39, 97-103.
  6. Choi, J.Y., Hong, S.W., and Chung, K.S. 2012. Selction of biogenic amine-reducing microorganisms from a traditional Korean-style fermented food, Cheonggukjang. Korean J. Food Sci. Technol. 44, 196-201. https://doi.org/10.9721/KJFST.2012.44.2.196
  7. Choi, U.K., Ji, W.D., Chung, H.C., Choi, D.H., and Chung, Y.G. 1997. Optimization for pigment production and antioxidative activity of the products by Bacillus subtilis DC-2. J. Korean Soc. Food Sci. Nutr. 26, 1036-1043.
  8. Han, G.H., Cho, T.Y., Yoo, M.S., Kim, C.S., Kim, J.M., Kim, H.A., Kim, M.O., Kim, S.C., Lee, S.A., Ko, Y.S., et al. 2007. Biogenic amines formation and content in fermented soybean paste (Cheonggukjang). Korean J. Food Sci. Technol. 39, 541-545.
  9. Jeon, S.B., Ryu, M.S., Kim, Y.S., Jo, S.W., Jeong, D.Y., and Uhm, T.B. 2013. Ioslation and identification of Bacillus strains with antagonistic properties against film-forming yeasts overgrown in low salted soybean pastes. Korean J. Microbiol. 49, 286-291. https://doi.org/10.7845/kjm.2013.3061
  10. Jeong, H.Y., Kim, T.H., Park, J.S., Kim, K.T., and Paik, H.D. 2003. Antioxidative and cholesterol-reducing activity of Bacillus polyfermenticus SCD. Korean J. Biotechnol. Bioeng. 18, 371-376.
  11. Jeong, S.J., Shin, M.J., Jeong, S.Y., Yang, H.J., and Jeong, D.Y. 2014. Characteristic analysis and production of short-ripened Korean traditional soy sauce added with rice bran. J. Korean. Soc. Food. Sci. Nutr. 43, 550-556. https://doi.org/10.3746/jkfn.2014.43.4.550
  12. Kang, D.W., Ryu, I.H., and Han, S.S. 2012. The isolation of Bacillus subtilis KYS-10 with antifungal activity against plant pathogens. Korean J. Pestic. Sci. 16, 178-186. https://doi.org/10.7585/kjps.2012.16.2.178
  13. Kim, Y.S., Cho, S.H., Jeong, D.Y., and Uhm, T.B. 2012. Isolation of biogenic amine-degrading strains of Bacillus subtilis and Bacillus amyloliquefaciens from traditionally fermented soybean products. Korean J. Microbiol. 48, 220-224. https://doi.org/10.7845/kjm.2012.042
  14. Kim, M.C. and Heo, M.S. 2005. Identification and cultural characterization of antioxidant producing bacteria isolated from the Jeju coasts. Kor. J. Life Science 15, 749-754. https://doi.org/10.5352/JLS.2005.15.5.749
  15. Kim, Y.S., Jeong, J.O., Cho, S.H., Jeong, D.Y., and Uhm, T.B. 2012a. Antimicrobial and biogenic amine-degrading activity of Bacillus licheniformis SCK B11 isolated from traditionally fermented red pepper paste. Korean J. Microbiol. 48, 163-170. https://doi.org/10.7845/kjm.2012.48.2.163
  16. Kim, Y.S., Lee, M.S., Yeom, J.H., Song, J.G., Lee, I.K., Yeo, W.H., and Yun, B.S. 2012b. Screening of antagonistic bacteria for biological control of ginseng root rot. Korean J. Mycol. 40, 44-48. https://doi.org/10.4489/KJM.2012.40.1.044
  17. Kim, S.K., Oh, S.J., Lee, S.J., Baek, Y.J., and Park, Y.H. 1994. Optimization conditions for the growth and bacteriocin production of Lactococcus sp. HY 449 using response surface methodology. Korean J. Appl. Microbiol. Biotechnol. 22, 522-530.
  18. Kim, B.S., Rhee, C.H., Hong, Y.A., Woo, C.J., Jang, C.M., Kim, Y.B., and Park, H.D. 2007. Changes of enzyme activity and physiological functionality of traditional Doenjang during fermentation using Bacillus sp. SP-KSW3. Korean J. Food Preserv. 14, 545-551.
  19. Kim, Y.S., Yun, S.H., Jeong, D.H., Hahn, K.S., and Uhm, T.B. 2010. Ioslation of Bacillus licheniformis producing antimicrobial agents against Bacillus cereus and its properties. Korean J. Microbiol. 46, 270-277.
  20. Kwak, H.J., Kim, J.Y., Lee, H.S., and Kim, S.M. 2014. Formation of biogenic amines by Lactobacillus plantarum isolated from Makgeolli. Korean J. Food Sci. Technol. 46, 438-445. https://doi.org/10.9721/KJFST.2014.46.4.438
  21. Lee, J.H., Chae, M.S., Choi, G.H., Lee, N.K., and Paik, H.D. 2009. Optimization of medium constituent for production of the antioxidant substances by Bacillus polyfermenticus SCD using response surface methodology. Food Sci. Biotechnol. 18, 959-964.
  22. Lee, G.D., Chang, H.G., and Kim, H.K. 1997. Antioxidative and nitrite-scavenging activities of edible Mushrooms. Korean J. Food Sci. Technol. 29, 432-436.
  23. Lee, H.J., Kim, J.Y., Lee, J.G., and Hong, S.S. 2014. Biological control of Lettuce sclerotinia rot by Bacillus subtilis GG95. Korean J. Mycol. 42, 225-230. https://doi.org/10.4489/KJM.2014.42.3.225
  24. Lee, J.H. and Kweon, K.T. 2012. Determination of harvest time and nominal origin from Cynanchi wilfordii radix. J. Korean Orient. Med. Soc. 33, 160-168.
  25. Lee, S.Y., Park, H.K., and Kim, M.H. 2010. Isolation and identification of wild yeasts from schizandra (Schizandra chinensis) for wine production and its characterization for physicochemical and sensory evaluations. J. Korean Soc. Food Sci. Nutr. 39, 1860-1866. https://doi.org/10.3746/jkfn.2010.39.12.1860
  26. Leadbetter, E.R. 1974. Order II. Cytophagales nomen novum, p. 99. In Buchanan, R.E. and Gibbons, N.E. (eds.), Bergey's Manual of Determinative Bacteriology, 8th ed. The Williams & Wilkins Co., Baltimore, Maryland, USA.
  27. Maijala, R. and Eerola, S. 1993. Contaminant lactic acid bacteria of dry sausages produce histamine and tyramine. Meat Sci. 35, 387-395. https://doi.org/10.1016/0309-1740(93)90043-H
  28. Oh, Y.S., Park, J.E., Oh, H.J., Kim, J.H., Oh, M.C., Oh, C.K., Oh, Y.J., and Lim, S.B. 2010. Isolation and characteristics of microorganisms producing extracellular enzymes from Jeju traditional fermented soybean paste (deonjang). J. Korean Soc. Food Sci. Nutr. 39, 47-53. https://doi.org/10.3746/jkfn.2010.39.1.047
  29. Park, J.H., Kim, J.H., and Kang, S.J. 2002. Isolation and antifungal activity of Bacillus ehimensis YJ-37 as antagonistic against vegetables damping-off fungi. Kor. J. Life Science 12, 200-207. https://doi.org/10.5352/JLS.2002.12.2.200
  30. Ra, K.S., Oh, S.H., Kim, J.M., and Suh, H.J. 2004. Isolation of fibrinolytic enzyme and ${\beta}$-glucosidase producing strains from Doenjang and optimum conditions of enzyme production. J. Korean Soc. Food Sci. Nutr. 33, 439-442. https://doi.org/10.3746/jkfn.2004.33.2.439
  31. Rhee, C.H., Lee, J.B., and Jang, S.M. 2000. Changes of microorganisms, enzyme activity and physiological functionality in the traditional Deonjang with various concentrations of Lentinus edodes during fermentation. J. Korean Soc. Agric. Chem. Biotechnol. 43, 277-284.
  32. Ryu, H.S., Shon, M.Y., Cho, S.J., Park, S.K., and Lee, S.W. 2007. Characterization of antibacterial substance-producing Bacillus subtilis isolated from traditional Doenjang. J. Korean Soc. Appl. Biol. Chem. 50, 87-94.
  33. Seki, T., Chung, C.K., Mikami, H., and Oshima, Y. 1978. Deoxyribonucleic acid homology and taxonomy of the genus Bacillus. Int. J. Syst. Bacteriol. 28, 182-189. https://doi.org/10.1099/00207713-28-2-182
  34. Teather, R. and Wood, P.J. 1982. Use of congo fed polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl. Environ. Microbiol. 43, 777-780.
  35. Vermelho, A.B., Meirelles, M.N.L., Lopes, A., Petinate, S.D.G., Chaia, A.A., and Branquinha, M.H. 1996. Detection of extracelluar proteases from microorganisms on agar plates. Mem. Inst. Oswaldo Cruz. 91, 755-760. https://doi.org/10.1590/S0074-02761996000600020
  36. Yang, E.J. and Chang, H.C. 2007. Characterization of bacteriocin-like substances produced by Bacillus subtilis MJP1. Korean J. Microbiol. Biotechnol. 35, 339-346.
  37. Yang, S.J., Lee, D.H., Park, H.M., Jung, H.K., Park, C.S., and Hong, J.H. 2014. Amylase activity and characterization of Bacillus subtilis CBD2 isolated from Doenjang. Korean J. Food Preserv. 21, 286-293. https://doi.org/10.11002/kjfp.2014.21.2.286
  38. Yi, J.H., Heo, N.K., Choi, B.G., Park, E.H., Kwun, S.Y., Kim, M.D., Hong, W.P., Yeo, S.H., and Baek, S.Y. 2014. Isolation of fibrinolytic yeasts Korean traditional fermented soybean. Korean J. Microbiol. Biotechnol. 42, 184-189. https://doi.org/10.4014/kjmb.1403.03006
  39. Yun, H.J., Lee, Y.J., Yeo, S.H., Park, H.Y., Park, H.D., and Baek, S.Y. 2013. Isolation and characterization of exopolysacchried producing lactic acid bacteria from Korean soy sauce and soybean paste. Korean J. Microbiol. Biotechnol. 41, 190-197. https://doi.org/10.4014/kjmb.1302.02004

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