• 제목/요약/키워드: Bacillus subtilis

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상동성 유전자재조합을 이용한 단백질분해효소 비생산 바실러스균주의 구축 (Construction of Pretense-defective Mutant of Bacillus subtilis by Homologous DNA Recombination)

  • Lee, Jin-Tae;An, Bong-Jeun
    • 한국식품저장유통학회지
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    • 제7권4호
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    • pp.414-417
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    • 2000
  • 단백질분해효소를 생산하지 않는 균주 B. subtilis MT-2의 염색체 DNA를 추출한 다음, B. subtilis AC819 균주에 상동성 유전자재조합을 이용하여 competent cell 형질전환을 시켰다. 얻어진 형질전환체를 B. subtilis HL-1이라고 명명하였으며, 그 표현형은 histidine 요구성, streptomycin 내성, tetracyclin 내성을 나타내면서 단백질 분해효소를 생산하지 않았다. 플라스미드 pUB110 을 이용한 B. subtilis HL-1의 protoplst 형질전환율은 B. subtilis MT-2의 형질전환율보다 높았다. 따라서 새로운 B. subtilis HL-1균주는 단백질분해효소의 형질전환과 내열성 protease 유전자클로닝에서 숙주로 사용하는데 유용하다.

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Properties of Bac W42, a Bacteriocin Produced by Bacillus subtilis W42 Isolated from Cheonggukjang

  • Kindoli, Salum;Lee, Hwang A;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • 제22권8호
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    • pp.1092-1100
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    • 2012
  • Ten Bacillus strains with antimicrobial activities were isolated from Cheonggukjang produced at different parts in Korea. They all inhibited Listeria monocytogenes ATCC 19111 and nine inhibited Bacillus cereus ATCC 14579. Four isolates (W42, H27, SKE 12, and K21) showing strong inhibiting activities were identified as B. subtilis. B. subtilis W42 was the most inhibiting strain. The antimicrobial activity of culture supernatant from B. subtilis W42 was destroyed completely by proteinase K treatment, indicating that a bacteriocin was the responsible agent. The bacteriocin, Bac W42, was most stable at pH 7 and stable between pH 3-6 and 8-9. Bac W42 was stable up to $80^{\circ}C$. BHI (brain heart infusion) and TSB (tryptic soy broth) were the best media for the activity (320 AU/ml) followed by LB (160 AU/ml). Bac W42 was partially purified by column chromatographies. The specific activity was increased from 1,151.2 AU/ml to 9,043.5 AU/ml and the final yield was 26.3%. Bac W42 was 5.4 kDa in size as determined by SDS-PAGE. Bac W42 showed bactericidal activity against L. monocytogenes ATCC 19111.

Bacillus subtilis subsp. krictiensis로부터 항진균물질 KRF-001의 생산을 위한 발효조건 및 돌연변이 연구 (Effect of Some Factors on the Production of an Antifungal Compound KRF-001 from Bacillus subtilis subsp. krictiensis)

  • 손광희;권혜경;복성해;이항우
    • 한국미생물·생명공학회지
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    • 제19권6호
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    • pp.614-618
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    • 1991
  • 토양세균 B.subtilis subsp. krictiensis로부터 항진균활성물질 KRF-001 복합체를 생산하기 위한 발효 조건을 조사하였다. Whey와 mannitol 등이 탄소원으로, CSL, corn gluten meal 그리고 polypeptone이 복합질소원으로 좋았다. 중온균인 생산균은 중성 pH조건에서 P.oryzae에 대한 bioactivity가 가장 높았으며, 0-10 DO의 낮은 용존산소 조건에서도 bioactivity가 유지되었다. 인산염의 농도가$ K_2HPO_4$, 0.047에서 0.097로 높아짐에 따라 1/2의 bioactivity 감소가 관찰되었다. 한편 UV 및 MNNG에 의한 돌연변이에 의해 우수균주 3개를 선별하였다.

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Bacterial Surface Display of $GFP_{UV}$ on Bacillus subtilis Spores

  • Kim, Jung-Hyung;Roh, Chang-Hyun;Lee, Chang-Won;Kyung, Do-Hyun;Choi, Soo-Keun;Jung, Heung-Chae;Pan, Jae-Gu;Kim, Byung-Gee
    • Journal of Microbiology and Biotechnology
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    • 제17권4호
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    • pp.677-680
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    • 2007
  • To analyze a cotG-based Bacillus subtilis spore display system directly, $GFP_{UV}$ was expressed on the surface of Bacillus subtilis spores. When $GFP_{UV}$ was fused to the C-terminal of the cotG structural gene and expressed, the existence of a $CotG-GFP_{UV}$ fusion protein on the B. subtilis spore was confirmed by flow cytometry confocal microscopic analysis. When the cotG anchoring motif was deleted, no fluorescence emission was observed under flow cytometry and confocal microscopic analysis from the purified spore, confirming the essential role of CotG as an anchoring motif. This $GFP_{UV}$ displaying spore might be used for another signaling application triggered by intracellular or extracellular stimuli.

Bacillus subtilis B-4228의 인삼 근부병 억제효과 (Control Effect of Bacillus subtilis B-4228 on Root Rot of Panax ginseng)

  • 이병대;밝훈
    • Journal of Ginseng Research
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    • 제28권1호
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    • pp.67-70
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    • 2004
  • 인삼 근부병의 생물학적 방제를 위해 인삼 재배토양으로부터 분리된 길항미생물 Bacillu subtilis B-4228을 사용하여 방제효과를 검정하였다. 공시 균주 B-4228과 C. destructans의 대치배양결과, C destructans의 균사가 비정상적인 형태로 팽창하였으며 강한 생장억제력을 보였다. C. destructans에 의해 오염된 이병토를 이용한 pot시험결과, 건전근율이 무처리구에서 20%였으나 공시 균주 B-4228의 처리구는 82%로 높게 나타났고, 이병율(개체 인삼에 대한 뿌리썩음 이병정도의 총합비율)은 무처리의 경우 50.4%였으나 공시 균주 B-4228의 처리구는 6.0%로 낮았다.

Phenolic Compounds Production, Enhancement and Its Antioxidant Activity of Blue Berry Powder with Bacillus subtilis Light Mediated Fermentation Compounds

  • Elumalai, Punniyakotti;Lim, Jeong-Muk;Mohan, Harshavardhan;Lee, Jeong-Ho;Oh, Byung-Taek
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 춘계학술발표회
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    • pp.66-66
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    • 2018
  • Light fermentation has been conducted under different light conditions such as normal dark light, white light, and light emitting diodes (LEDs) various color (blue, green, red, white on blueberry powder with fermenting bacteria Bacillus subtilis (B2). The bacteria B2 was isolated and identified by 16S rRNA sequencing method. RYRP biologically converted to secondary metabolites through light fermentation in the presence of Bacillus subtilis, the bacteria actively involved in bioconversion process. LEDs fermentation to enhance the production of phenolic content while comparing to normal dark and white light. Among the different color LEDs, blue LEDs mediated fermentation showed higher amount of total phenolic and flavonoid content. Then blue LEDs mediated fermented compound were characterized by FTIR and GC-MS, subsequently the compound was analyzed antioxidant activity tests and the antioxidant activity exhibited higher. This is the first study to demonstrate that B. subtilis-LEDs mediated fermentation is useful for facilitating phenolic compound production and enhancing antioxidant activity, which may have greater application fermentation fields.

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Cytotoxicity of Listeriolysin O Produced by Membrane-Encapsulated Bacillus subtilis on Leukemia Cells

  • Stachowiak, R.;Granicka, L.H.;Wisniewski, J.;Lyzniak, M.;Kawiak, J.;Bielecki, J.
    • Journal of Microbiology and Biotechnology
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    • 제21권11호
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    • pp.1193-1198
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    • 2011
  • Encapsulation of biological material in the permiselective membrane allows to construct a system separating cells from their products, which may find biotechnological as well as biomedical applications in biological processes regulation. Application of a permiselective membrane allows avoiding an attack of the implanted microorganisms on the host. Our aim was to evaluate the performance of Bacillus subtilis encapsulated in an elaborate membrane system producing listeriolysin O, a cytolysin from Listeria monocytogenes, with chosen eukaryotic cells for future application in anticancer treatment. The system of encapsulating in membrane live Bacillus subtilis BR1-S secreting listeriolysin O was proven to exert the effective cytotoxic activity on eukaryotic cells. Interestingly, listeriolysin O showed selective cytotoxic activity on eukaryotic cells: more human leukemia Jurkat T cells were killed than human chronic lymphocytic B cells leukemia at similar conditions in vitro. This system of encapsulated B. subtilis, continuously releasing bacterial products, may affect selectively different types of cells and may have future application in local anticancer treatment.

Antifungical Activity of Autochthonous Bacillus subtilis Isolated from Prosopis juliflora against Phytopathogenic Fungi

  • Abdelmoteleb, Ali;Troncoso-Rojas, Rosalba;Gonzalez-Soto, Tania;Gonzalez-Mendoza, Daniel
    • Mycobiology
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    • 제45권4호
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    • pp.385-391
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    • 2017
  • The ability of Bacillus subtilis, strain ALICA to produce three mycolytic enzymes (chitinase, ${\beta}$-1,3-glucanase, and protease), was carried out by the chemical standard methods. Bacillus subtilis ALICA was screened based on their antifungal activity in dual plate assay and cell-free culture filtrate (25%) against five different phytopathogenic fungi Alternaria alternata, Macrophomina sp., Colletotrichum gloeosporioides, Botrytis cinerea, and Sclerotium rolfesii. The B. subtilis ALICA detected positive for chitinase, ${\beta}$-1,3-glucanase and protease enzymes. Fungal growth inhibition by both strain ALICA and its cell-free culture filtrate ranged from 51.36% to 86.3% and 38.43% to 68.6%, respectively. Moreover, hyphal morphological changes like damage, broken, swelling, distortions abnormal morphology were observed. Genes expression of protease, ${\beta}$-1,3-glucanase, and lipopeptides (subtilosin and subtilisin) were confirmed their presence in the supernatant of strain ALICA. Our findings indicated that strain ALICA provided a broad spectrum of antifungal activities against various phytopathogenic fungi and may be a potential effective alternative to chemical fungicides.

Bacillus subtilis BB-1으로부터 나토키나아제 유전자 크로닝 및 대량발현 (Cloning and High Expression of Nattokinase Gene from Bacillus subtilis BB-1)

  • 이영훈;이성호;박기훈;최영주;정영기;갈상완
    • 생명과학회지
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    • 제16권2호
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    • pp.274-281
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    • 2006
  • 흑두청국으로부터 분리된 혈전용해력이 우수한 Bacillus subtilis BB-1(KFCC 11344P)으로부터 혈전용해효소 유전자를 PCR법에 의해 크로닝하였고 이를 BCF-1으로 명명하였다. BCF-1의 DNA 염기서열결정 결과 1,145 bp 크기의 혈전용해 효소로, 일본의 natto로부터 분리된 nattokinase 유전자와 99%의 상동성을 보임을 확인하였다. 혈전용해효소 유전자의 발현을 위하여 Bacillus 발현계인 Bacillus-E. coli의 shuttle vector인 pEB vector에 크로닝 하고 host로서 B. subtilis 168에 형질전환시켜 대량 발현시켰다. 생산된 혈전용해효소의 최 적활성 pH와 온도는 7.0과 $35^{\circ}C$로 확인되었다, 기질에 대한 분해양상을 조사한 결과 fibrin에서만 특이적으로 강한 분해가 일어났으며, skim milk에서 아주 약한 분해능을 보였으나 blood agar, gelatin, casein에서는 전혀 분해능을 보이지 않았다. 특히 blood agar plate에서 분해능이 없는 것으로 보아 혈액 내에서의 적혈구 파괴현상과 같은 부작용에 대한 위험을 배제할 수 있을 것으로 사료된다. BCF-1에 의해 생산된 혈전용해효소는 fibrin 특이적으로 활성을 나타냄을 확인할 수 있으며, 이는 임상적이나 산업적으로 적용하였을 때 부작용에 대한 위험적인 문제는 배제될 수 있으리라 생각된다.

Edwardsiella tarda의 특이 Bacteriophage와 Bacillus subtilis KM-1혼합액이 Edwardsiella tarda 에 미치는 항균효과 (Mixture of Edwardsiella tarda specific Bacteriophage and Bacillus subtilis KM-1enhanced bactericidal activity against Edwardsiella tarda)

  • 백민석;황요셉;최상훈
    • 한국어병학회지
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    • 제26권3호
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    • pp.185-191
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    • 2013
  • 본 연구는 E. tarda에 대한 특이박테리오phage와 생균제의 일종인 B. subtilis KM-1의 혼합제가 E. tarda에 대한 항균활성에 미치는 효과를 알아보고자 수행되었다. 가장 효과적인 혼합제의 항균활성을 보인 phage의 적정 수는 B. subtilis 가 $2{\times}10^5$ CFU/ml의 농도로 존재할 때 배양 36시간째 $2{\times}10^5$ PFU/ml로 나타났다. B. subtilis의 경우 $2{\times}10^5$ PFU/ml의 phage가 존재할 때 첨가량에 비례하는 항균활성을 보여주었다. phage와 B. subtilis의 혼합투여는 phage나 B. subtilis 각각을 투여한 그룹보다 훨씬 높은 항균활성을 나타냈다. 본 연구의 결과는 수산양식에서 대두되고 있는 양식어류의 E. tarda 감염성 질병 치료를 위한 항생제 대체제로서의 사료첨가제 개발 가능성을 제시하고 있다.