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Bacillus subtilis MJP1이 생산하는 Bacteriocin-Like Substances
Characterization of Bacteriocin-Like Substances Produced by Bacillus subtilis MJP1

양은주   (조선대학교 식품영양학과UU0001180  ); 장해춘   (조선대학교 식품영양학과UU0001180  );
  • 초록

    메주로부터 항곰팡이 활성을 보이는 균주 1종과 이에 감수성을 나타내는 곰팡이 3종을 분리하였다. 분리된 균주는 형태학적, 생화학적 특성 조사와 16S rRNA 염기서열 결정을 통한 균주 동정결과 Bacillus subtilis MJP1으로 명명하였고, 3종의 곰팡이는 ITS-5.85 rRNA염기서열 분석을 통하여 Aspergillus petrakii PF-1, A. ochraceus PF-2, 그리고 A. nidulans PF-3로 명명하였다 B. subtilis MJP1은 곰팡이 에 대한 강한 저해활성 뿐만 아니라 Candida 속 효모들과 그람 양성균에 대한 넓은 범위의 저해활성을 나타내었다. B. subtilis MJP1의 생육에 따른 항균 활성을 측정한 결과 항진균 활성은 배양 16시간부터 최대 활성(3,200 AU/ml)을 나타내어 균이 사멸기에 접어든 후에도 활성을 그대로 유지한 반면, 항세균 활성은 대수기 중반인 12시간부터 25시간까지 가장 높은 활성 (1,600 AU/ml)을 보이다가 72시간 이후에는 활성을 상실하였다. pH 안정성 실험에서 항진균 활성과 항세균 활성 모두 pH $6{\sim}10$ 구간에서 비교적 안정한 결과를 보였으나, 열처리 실험에서 항진균 활성은 영향을 받지 않은 반면, 항세균 활성은 $30^{\circ}C$ 이상의 온도에서는 불안정한 결과를 보였다. 각종 효소에 대한 안정성 실험에서 항진균 활성은 어떠한 효소에도 영향을 받지 않았으나, 항세균 활성은 단백분해효소 처리 후에 활성이 실활 됨으로써 항균 물질이 단백질성 물질임을 추정하였다. $C_{18}$ Sep-Pak column으로 부분 정제한 항균 물질이 항진균 활성과 항세균 활성을 나타내므로 소수성을 가지는 물질임을 알 수 있으며, Tricine-SDS-PAGE및 direct detection을 통하여 항진균 물질의 분자량은 약 2.4 kDa 정도이며, 항세균 물질의 분자량은 약 4.5 kDa으로_ 확인되었다. 따라서 B. subtilis MJP1은 항진균 활성과 항세균 활성을 가진 bacteriocin-like substances를 생산함을 알 수 있고 이와 같은 새로운 항미생물 물질은 천연 식품보존제 및 사료보존제 뿐만 아니라 항생제 대체 의약품으로도 활용이 기대되며, 이를 위하여 향후 이 물질들의 보다 정확한 구조 및 특성 규명 등의 연구가 필요하다.


    The MJP1 bacterial strain, which possesses antifungal activity, was isolated from meju and identified as Bacillus subtilis based on its morphological and biochemical properties, as well as its 16S rRNA sequence. Antimicrobial activity was found against various species of Gram-positive bacteria, yeasts, and molds, including food-spoilage microorganisms. The antifungal activity was found to be stable after heat and proteolytic enzyme treatment, and in the pH range of $6.0{\sim}10.0$ . The antibacterial activity was stable in the pH range of $6.0{\sim}10.0$ , but about 50% of the activity was lost after 24 hr at $30^{\circ}C$ . The antibacterial compound was also inactivated by proteolytic enzyme treatment, indicating its proteinaceous nature. The apparent molecular masses of the partially purified antifungal and antibacterial compounds, as indicated by using the direct detection method in Tricine-SDS-PAGE, were approximately 2.4 kDa and 4.5 kDa, respectively. These studies suggest that B. subtilis MJP1 produces two bacteriocin-like substances with antifungal and antibacterial activities.


  • 주제어

    Bacillus subtilis .   bacteriocin-like substance .   antifungal activity .   antibacterial activity.  

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  • 이 논문을 인용한 문헌 (13)

    1. Jung, Ji-Hye ; Chang, Hae-Choon 2009. "Antifungal Activity of Bacillus polyfermenticus CJ6 Isolated from Meju" 한국식품영양과학회지 = Journal of the Korean Society of Food Science and Nutrition, 38(4): 509~516     
    2. Chung, Jae Hyuk ; Bae, Yun-Sook ; Kim, Yeon-Joo ; Lee, Jong-Hoon 2010. "Characteristics of Bacteriocin Produced by a Lactobacillus plantarum Strain Isolated from Kimchi" 한국미생물·생명공학회지 = Korean journal of microbiology and biotechnology, 38(4): 481~485     
    3. Kim, Yong-Sang ; Yun, Suk-Hyun ; Jeong, Do-Yeon ; Hahn, Kum-Su ; Uhm, Tai-Boong 2010. "Isolation of Bacillus licheniformis Producing Antimicrobial Agents against Bacillus cereus and Its Properties" Korean journal of microbiology = 미생물학회지, 46(3): 270~277     
    4. Kwon, Gun-Hee ; Lee, Hwang-A ; Kim, Jeong-Hwan 2010. "A Bacteriocin of 5-kDa in Size Secreted by Bacillus subtilis 168" 한국미생물·생명공학회지 = Korean journal of microbiology and biotechnology, 38(2): 163~167     
    5. Yim, Eun-Jung ; Yang, Eun-Ju ; Chang, Hae-Choon 2010. "Purification and Characterization of Antibacterial Compound Produced by Bacillus subtilis MJP1" 한국미생물·생명공학회지 = Korean journal of microbiology and biotechnology, 38(1): 84~92     
    6. Lee, Seung-Gyu ; Han, Ki-Sung ; Jeong, Seok-Geun ; Oh, Mi-Hwa ; Jang, Ae-Ra ; Kim, Dong-Hun ; Bae, In-Hyu ; Ham, Jun-Sang 2010. "A Study on the Sensory Characteristic of Yogurt and Antimicrobial Activity of Lactobacillus plantarum LHC52 Isolated from Kimchi" Korean journal for food science of animal resources = 한국축산식품학회지, 30(2): 328~335     
    7. Jung, Ji-Hye ; Chang, Hae-Choon 2011. "Characterization of Antibacterial Compounds from Bacillus polyfermenticus CJ6 and Its Growth Inhibition Effect on Food-Borne Pathogens" 한국식품영양과학회지 = Journal of the Korean Society of Food Science and Nutrition, 40(6): 903~911     
    8. Chang, Mi ; Moon, Song Hee ; Chang, Hae Choon 2012. "Isolation of Bacillus velezensis SSH100-10 with Antifungal Activity from Korean Traditional Soysauce and Characterization of Its Antifungal Compounds" 한국식품저장유통학회지 = Korean journal of food preservation, 19(5): 757~766     
    9. Park, Sungyong ; Yoo, Jincheol ; Seong, Chinam ; Cho, Seungsik 2013. "Isolation and Characterization of Bacillus subtilis MP56 with Antimicrobial Activity against MDR (Multi Drug Resistant) Strains" Korean journal of microbiology = 미생물학회지, 49(1): 90~94     
    10. Lee, Jun-Ki ; Jo, Hyeon-Ju ; Yoon, Jin-A ; Chung, Kang-Hyun ; Song, Byeong Chun ; Kim, Kyoung Im ; An, Jeung Hee 2014. "Isolation and Identification of Microorganisms with Antimicrobial Activity in Makgeolli of Different Kinds Koji and Nuruk" 한국식품영양과학회지 = Journal of the Korean Society of Food Science and Nutrition, 43(4): 577~583     
    11. Kim, Hyo Ju ; Sung, Hea Mi ; Shin, Hyun-Kyung ; Kim, Ki Myong ; Yang, Eun Ju 2014. "Characterization of Probiotic and Functional Properties of Lactobacillus curvatus ML17, a Mukeunji Starter" 한국식품영양과학회지 = Journal of the Korean Society of Food Science and Nutrition, 43(7): 1009~1016     
    12. Lim, Joung-Soo ; Cho, Sung-Back ; Hwang, Ok-Hwa ; Yang, Seung-Hak 2014. "Isolation and Characterization of Phosphorus Accumulating Microorganisms under Liquid Fertilization of Swine Slurry" 한국축산시설환경학회지 = Journal of animal environmental science, 20(2): 77~84     
    13. Lim, Joung-Soo ; Han, Deug-Woo ; Lee, Sang-Ryong ; Hwang, Ok-Hwa ; Kwag, Jung-Hoon ; Cho, Sung-Back 2015. "A Bacterial Strain Identified as Bacillus licheniformis using Vitek 2 Effectively Reduced NH3 Emission from Swine Manure" 한국축산시설환경학회지 = Journal of animal environmental science, 21(3): 83~92     

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