한국식품영양과학회지 (Journal of the Korean Society of Food Science and Nutrition)

  • 제39권1호
  • /
  • Pages.47-53
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  • 2010
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  • 1226-3311(pISSN)
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  • 2288-5978(eISSN)
한국식품영양과학회 (The Korean Society of Food Science and Nutrition)
권호 표지
DOI QR코드

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제주전통된장으로부터 세포외효소 분비능이 우수한 미생물의 분리 및 특성

Isolation and Characteristics of Microorganisms Producing Extracellular Enzymes from Jeju Traditional Fermented Soybean Paste (Doenjang)

  • 오유성 (제주대학교 생명과학기술혁신센터) ;
  • 박지은 (제주대학교 생명과학기술혁신센터) ;
  • 오현정 (제주대학교 생명과학기술혁신센터) ;
  • 김정현 (제주관광대학 관광외식조리계열) ;
  • 오명철 (제주산업정보대학 관광호텔조리과) ;
  • 오창경 (제주산업정보대학 관광호텔조리과) ;
  • 오영주 (제주한라대학 호텔조리과) ;
  • 임상빈 (제주대학교 생명과학기술혁신센터)
  • Oh, You-Sung (Biotechnology Regional Innovation Center, Jeju National University) ;
  • Park, Ji-Eun (Biotechnology Regional Innovation Center, Jeju National University) ;
  • Oh, Hyun-Jeong (Biotechnology Regional Innovation Center, Jeju National University) ;
  • Kim, Jung-Hyon (Dept. of Tourism & Food Service Cuisine, Cheju Tourism College) ;
  • Oh, Myung-Cheol (Dept. of Tourism Hotel Culinary Arts, Jeju College of Technology) ;
  • Oh, Chang-Kyung (Dept. of Tourism Hotel Culinary Arts, Jeju College of Technology) ;
  • Oh, Young-Ju (Dept. of Hotel Culinary Arts, Jeju Halla College) ;
  • Lim, Sang-Bin (Biotechnology Regional Innovation Center, Jeju National University)
  • 발행 : 2010.01.30
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초록

제주 전통된장으로부터 세포외효소(protease, fibrinolytic enzyme, amylase, cellulase, lipase) 분비능이 우수한 세균을 분리한 후 16S rRNA 유전자 분석과 생리적 특성을 분석하여 균주를 확인하고자 하였다. Protease 분비능은 JR14, JR19, JR25, JR32, JR38, JR47과 JR64가 표준균주인 Bacillus subtilis KCCM12027보다 활성이 높았다. Amylase 분비능은 JR6, JR25, JR38, JR56, JR81에서 나타난 반면 표준균주인 KCCM12027에서는 나타나지 않았다. Cellulase 분비능은 JR6, JR14, JR48과 JR65가 다른 분리균주 또는 표준균주보다 높았으며, lipase 분비능은 JR14와 JR48이 높았다. 혈전용해 활성은 positive control인 plasmin의 용해 영역에 비하여 JR19가 192%로 가장 높았고, hemolysis 활성도 높았다. 혈전용해능이 있는 균주인 JR19, JR32, JR47, JR64의 배양액을 zymography한 결과, 25～75 kDa 사이에서 4～5개의 밴드가 확인되었다. 된장으로부터 분리한 균주들의 16s rRNA 유전자 염기서열을 분석한 결과 모두 Bacillus species와 99%의 상동성을 나타내었으며, 혈전용해능이 가장 우수한 JR19는 B. stratosphericus $41KF2a^T$와 거의 일치하였다.

Bacteria strains with high activities of extracellular enzymes (protease, fibrinolytic enzyme, amylase, cellulase, and lipase) were isolated from Jeju traditional fermented soybean paste (Doenjang), and characterized by 16S rRNA gene sequence analysis and physiological properties. Protease activities were higher in JR14, JR19, JR25, JR32, JR38, JR47, and JR64 than Bacillus subtilis KCCM 12027 (standard strain). Amylase activities were shown in JR6, JR25, JR38, JR56 and JR81, while not in KCCM12027. Cellulase activities were higher in JR6, JR14, JR48, and JR65 than those of other isolated strains and KCCM 12027 whereas lipase activities were the higher in JR-14 and JR-48. Thrombolytic activity in JR19 with high hemolysis activity were 192% compared with that of plasmin as a positive control. Zymogram analysis indicated that the thrombolytic active strains had 4~5 bands in the molecular weight range of 25~75 kDa. Gene sequence analysis of rRNA revealed that the isolated stains had 99% homology with Bacillus species, and the thrombolytic active stain JR19 was B. stratosphericus $41KF2a^T$.

키워드

참고문헌

  1. Yoshikatsu M, Mitsuo Y. 2008. Traditional healthful fermented products of Japan. J Ind Microbiol Biotechnol 35: 791-798. https://doi.org/10.1007/s10295-008-0362-5
  2. Susumu M, Hitomi Y, Toshiko K, Tomoyuki S, Ryohei FT, Ikuko N. 2008. Immunomodulatory effect of halophilic lactic acid bacterium Tetragenococcus halophilus Th221 from soy sauce moromi grown in high-salt medium. Int J Food Microbiol 121: 245-252. https://doi.org/10.1016/j.ijfoodmicro.2007.10.011
  3. Kwon HY, Ryu HY, Kwon CS, Lee SH, Sohn HY. 2007. Optimization of culture conditions of Bacillus pumilis JB-1 for Chungkookjang fermentation in soybean boiling-waste liquor medium. Kor J Microbiol Biotechnol 35: 304-309.
  4. Yoo SK, Cho WH, Kang SM, Lee SH. 1999. Isolation and identification of microorganisms in Korean traditional soybean paste and soybean sauce. Kor J Appl Microbiol Biotechnol 27: 113-117.
  5. Feldeisen SE, Tucker KL. 2007. Nutritional strategies in the prevention and treatment of metabolic syndrome. Appl Physiol Nutr Metab 32: 46-60. https://doi.org/10.1139/H06-101
  6. Phromraksa P, Nagano H, Boonmars T, Kamboonruang C. 2008. Identification of proteolytic bacteria from Thai traditional fermented food and their allergenic reducing potentials. J Food Sci 73: 189-195. https://doi.org/10.1111/j.1750-3841.2008.00721.x
  7. Kim SB, Lee DW, Cheigh CI, Cheo EA, Lee SJ, Hong YH, Choi HJ, Pyun YR. 2006. Purification and characterization of a fibrinolytic subtilisin-like protease of Bacillus subtilis TP-6 from an Indonesian fermented soybean, Tempeh. J Ind Microbiol Biotechnol 33: 436-444. https://doi.org/10.1007/s10295-006-0085-4
  8. Deepak V, Kalishwaralal K, Ramkumarpandian S, Venkatesh BS, Senthilkumar SR. 2008. Optimization of media composition for nattokinase production by Bacillus subtilis using response surface methodology. Biores Technol 99: 8170-8174. https://doi.org/10.1016/j.biortech.2008.03.018
  9. Ro YM. 1998. How important is the mode of thrombolytic agents administration for optimal thrombolysis?－comments on the results of double bolus urokinase regimen in thrombolysis in myocardial infarction in Korea (TIMIKO) multicenter trial. Kor Circul J 28: 1661-1663. https://doi.org/10.4070/kcj.1998.28.10.1661
  10. Omura K, Hitosugi M, Zhu X, Ikeda M, Maeda H, Tokudome S. 2005. A new derived protein from Bacillus subtilis natto with both antithrombotic and fibrinolytic effects. J Pharmacol Sci 99: 247-251. https://doi.org/10.1254/jphs.FP0050408
  11. Ra KS, Oh SH, Kim JM, Suh HJ. 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
  12. Ryu BH. 2003. Development of functional Doenjang for antioxidative and fibrinolytic activity. Kor J Life Sci 13: 559-568. https://doi.org/10.5352/JLS.2003.13.5.559
  13. Hyun KW, Lee JS, Ham JH, Choi SY. 2005. Isolation and identification of microorganism with potent fibrinolytic activity from Korean traditional Doenjang. Kor J Microbiol Biotechnol 33: 24-28.
  14. Shin GM. 2004. Studies on quality characteristics of the traditional Doenjangs in Jeju. MS Thesis. Jeju National University, Jeju, Korea.
  15. Hyun GT, Koh SH, Chun CB, Lee CH. 2004. Studies on the contents of isoflavone in traditional soybean paste and soybeans in Jeju-do. Report of J.I.H.E. 15: 45-66.
  16. Astrup T, Mullerttz S. 1952. The fibrin method for estimating of fibrinolytic activity. Arch Biochem Biophys 40: 346-351. https://doi.org/10.1016/0003-9861(52)90121-5
  17. Oh HJ, Lim JH, Lee JY, Jeon BS, Kang HY, Oh YS, Oh YJ, Lim SB. 2009. Quality characteristics of Jeju traditional Doenjang. Kor J Cul Res 15: 298-308.
  18. Choi NS, Chung DM, Han YJ, Kim SH, Song JJ. 2009. Purification and Characterization of a subtilisin D5, a fibrinolytic enzyme of Bacillus amyloliquefaciens DJ-5 isolated from Doenjang. Korean J Food Sci Technol 18: 500-505.
  19. Kim DH, Song HP, Kim JO, Byun MW. 2004. A correlation between fibrinolytic activity and microflora in Korean fermented soybean products. J Korean Soc Food Sci Nutr 33: 41-46. https://doi.org/10.3746/jkfn.2004.33.1.041
  20. Lee SK, Bae DH, Kwon TJ, Lee SB, Lee HH, Park JH, Heo S, Johnson MG. 2001. Purification and characterization of a fibrinolytic enzyme from Bacillus sp. KDO-13 isolated from soybean paste. J Microbiol Biotechnol 11: 845-852.
  21. Wang CT, Ji BP, Li B, Nout R, Li PL, Ji H, Chen LF. 2006. Purification and characterization of a fibrinolytic enzyme of Bacillus subtilis DC33, isolated from Chinese traditional Douchi. J Ind Microbiol Biotechnol 33: 750-758. https://doi.org/10.1007/s10295-006-0111-6
  22. Gabriela S, Jaroslava S. 2008. Review of surfactin chemical properties and potential applications. Cent Eur J Med 3: 123-133. https://doi.org/10.2478/s11536-008-0002-5
  23. Lijnen HR. 2002. Matrix metalloproteinases and cellular fibrinolytic activity. Biochem (Moscow) 67: 92-98. https://doi.org/10.1023/A:1013908332232
  24. Jeong SJ, Kwon GH, Chun J, Kim JS, Park CS, Kwon DY, Kim JH. 2007. Cloning of fibrinolytic enzyme gen from Bacillus subtilis isolated from Cheonggukjang and its expression in protease-deficient Bacillus subtilis strains. J Microbiol Biotechnol 17: 1018-1023.

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