• 한국미생물·생명공학회지
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• 제17권2호
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• pp.160-164
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• 1989

알카리성 Bacillus sp. AL-8의 알카리성 amylase 유전자를 포함하는 5.7Kb의 EcoRI 단편을 pUB 110을 vector로 하여 amylase를 생산하지 못하는 B. subtilis sta-1에서 발현시켰다. 재조합 plasmid pMB802와 pMB809는 숙주세포인 B. subtilis에서 매우 안정하게 유지되었으며 amylase 생산이 공여균 주에서 보다 1.8배 증가하였다. 형질전환주에서 생산된 amylase는 공여균주와 같은 효소적 성질을 나타내었다. 5.7Kb 단편을 E. coli에 subcloning한 결과 3.7Kb의 EcoRI 단편에 알카리성 amylase 유전자가 존재하였다.

• 한국식품위생안전성학회지
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• 제15권2호
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• pp.122-127
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• 2000

아플라톡신 생성균의 생육을 억제할 수 있는 미생물을 분리동정하고 진공동결 건조한 대사산물로부터 길항물질을 분리하였다. 대두로부터 아플라톡신 생육을 저해하는 길항균을 분리하였으며, 그 균은 Bacillus subtilis로 동정되었다. Bacillus subtilis는 아플라톡신 생성균에 대한 길항물질을 생산하였으며 MeOH추출, 실리카겔 흡착 크로마토그래피법, Sephadex LH-20 크로마토그래피법으로 길항물질을 분리하였다. 아플라톡신 생성균인 Asp. flavus와 Asp. parasiticus의 생육은 길항미생물인 Bacillus subtilis가 생산하는 길항물질에 의해 강하게 저해되었으며 이는 아플라톡신 오염방지를 위하여 효과적인 생물학적 방법일 것이라 기대된다.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1372-1375
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• 2008

The aprE2 gene from Bacillus subtilis CH3-5 was expressed in Bacillus licheniformis ATCC 10716 using a Bacillus-Escherichai coli shuttle vector, pHY300PLK. The fibrinolytic activity of transformant (TF) increased significantly compared to B. licheniformis 10716 control cell. During the 100 hr incubation in Luria-Bertaini broth at $37^{\circ}C$, fibrinolytic activity of B. licheniformis TF increased rapidly at the late growth stage, after 52 hr of incubation, which was confirmed by zymography using a fibrin gel. pHY3-5 was stably maintained in B. licheniformis without tetracycline (Tc) in the media, 60.9% of cells still maintained pHY3-5 after 100 hr of cultivation.

• 식물병연구
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• 제12권2호
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• pp.85-90
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• 2006

차나무 겹퉁근무늬병을 일으키는 Pestalotiopsis longiseta에 대하여 강력한 길항능력을 나타내는 Bacillus subtilis BD0310 균주의 대량배양을 위한 배양조건과 P. longiseta에 대한 항균활성을 증대시킬 수 있는 탄소원과 질소원을 선발하였다. B. subtilis BD0310 균주를 대량배양하기 위한 최적온도 및 시간은 $30^{\circ}C$, 24 시간인 것으로 확인되었으며, 초기 최적 pH 는 7 로 확인되었다. B. subtilis BD03l0 균주를 대량으로 배양할 경우 P. longiseta에 대한 항균활성을 가장 높게 증가시키는 탄소원을 선발하기 위하여 fructose, galactose, glucose, glycerol, inositol, lactose, maltose, sorbitol, starch 등 9 가지 탄소원을 사용하여 조사한 결과 maltose와 inositol이 가장 효율적인 탄소원으로 선발되었으며, casein, tryptone, malt extract, yeast extract, $(NH_4)_2SO_4$등 5 가지 질소원 중에서 yeast extract와 tryptone이 가장 효율적인 질소원으로 선발되었다. 이러 한 결과들은 길항세균 B. subtilis BD0310를 차나무 겹둥근무늬병 방제용 미생물제제로 개발하기 위한 대량배양생산 공정을 확립하는데 기여할 것으로 전망된다.

• 환경생물
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• 제22권4호
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• pp.487-493
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• 2004

To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA sequencing. It has an antifungal activity against 9 plant pathogenic fungi in a paper disc assay. The antifungal activity- deficient mutant, B. subtilis mLTD was induced at a 5 kGy dose of $^{60}Co$ gamma radiation. Using the two-dimensional electrophoresis and the matrix assisted laser desorption ionization time-of-flight mass spectrometry, the comparison analysis of proteins between the wild and mutant were performed. A major intracellular serine proteinase IspA (MW: 32.5 kDa), a NAD (P) H dehydrogenase (MW: 20.0 kDa), and a stage II sporulation protein AA, SpoIIAA (MW: 14.3kDa) were detected only in the B. subtilis LTD. These results suggested that the functions of these proteins found only in the B. subtilis LTD could. be closely related to the antifungal activity against plant pathogenic fungi.

• 한국의류산업학회지
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• 제8권2호
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• pp.239-244
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• 2006

For studies of fibrinolytic enzyme strain K-54 was isolated from the Korean traditional food chungkook-jang. Isolated strains K-54 was identified as Bacillus subtilis. The molecular weight of fibrinolytic enzyme from B. subtilis K-54 was 27 kDa. Optimum temperature for fibrinolytic enzyme of B. subtilis K-54 was $50-70^{\circ}C$ and optimum pH for producing the enzyme of this strain was ranging from 8 to 12. Also, it was found out enzyme activity was completely inhibited by 1mM PMSF. The result indicated this enzyme was thermo-stable alkaline serine protease with strong fibrinolytic activity. The wool and silk were treated with protease of B. subtilis K-54. As a result, the property of dyeing of wool fabrics was increased. By the increasing of treatment time became smoothened. But the change of mechanical properties were not changed.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.128-137
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• 2023

Transcriptome analysis revealed that the sinR gene encoding a transition-state regulator of Bacillus pumilus, genetically close to B. subtilis, was expressed at high levels during growth. The sinR gene is the second gene of the sinIR operon consisting of three promoters and two structural genes in B. subtilis. This study used the sinIR promoter of B. subtilis DB104 to construct a recombinant protein expression system. First, the expression ability depending on the number of sinIR promoter was investigated using enhanced green fluorescent protein (eGFP). The expression level of eGFP was slightly higher when using two promoters (Psin2) than using original promoters. The Psin2 promoter was further engineered by modifying the repressor binding site and -35 and -10 regions. Shine-Dalgarno (SD) sequence of the sinI gene was modified to the consensus sequence. Finally, combining the engineered Psin2 promoter with the modified SD sequence increased the expression level of eGFP by about 13.4-fold over the original promoter. Our results suggest that the optimized sinIR promoter could be used as a novel tool for recombinant protein expression in B. subtilis.

• 한국미생물·생명공학회지
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• 제26권2호
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• pp.122-129
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• 1998

최근 생물방제균으로 주목받고 있는 Enterobacter cloacae의 방제기작이 이 균에 의해 토양내에서 생산된 휘발성 ammonia이며 ammonia의 생산에는urease가 관계한다는 보고를 근거로 하여, 항생물질 생산성 균주로 선발된 우수한 길항균주에 암모니아 생성능, 즉 urease 유전자를 유전적으로 부가함으로써 항진균성 길항물질 생산과 암모니아 생산이 동시에 이루어 질 수 있는 새로운 다기능의 생물방제균을 유전적으로 육종하고자 하였다. 저병해 인삼경작지로부터 식물근부균 Fusarium solani의 생육을 강하게 억제하는 길항세균 한 균주 SH14균주를 분리, 선발하였으며, 분리된 균주를 동정한 결과 Bacillus subtilis이거나 그 근연종으로 추정되었다. 억제기작 실험을 통해 길항균주 B. subtilis SH14에 의해 생산되는 항진균성 길항물질은 외막가수분해효소와 같은 고분자 물질이 아니라 열에 안정한 저분자의 항생물질임을 알 수 있었다. 한편 ammonia 생산을 위한 urease의 유전자는 urease 생산력이 강력한 호알칼리성 Bacillus pasteurii의 urease 생산유전자를 E. coli-Bacillus shuttle vector인 pEB203에 subcloning하였고, 이어서 pGU 366으로 명명된 이 recombinant plasmid를 선발된 항진균성 길항균주 B. subtilis SH14에 PEG-induced protoplast transformation 방법으로 도입, 발현시켰으며, 최적조건을 조사하여 90분간의 lysozyme 처리과정 후 1.5 $\mu\textrm{g}$/$m\ell$의 DNA와 40% PEG4000의 첨가로 약 6.5$\times$$10^{-4}$의 형질전환율을 얻을 수 있었다. 아울러 암모니아 생성능이 부가된 생물방제균 B. subtilis SH14(pGU366)에 의해 식물근부균 F. solani에 대한 생육억제력이 증가되는지 여부를 억제거리 측정법과 균체중량법을 통해 확인한 결과 urease 유전자가 도입된 형질전환체 B. subtilis SH14(pGU366)의 근부균 생육억제능이 각각 36.7%, 44.0%정도로 숙주균주인 B. subtilis SH14에 비해 근부균 생육억제능을 보다 강하게 나타내었음을 알 수 있었다. 따라서 항진균성 항생물질 생산성 생물방제균 B. subtilis SH14에 외부의 urease유전자를 도입하여 ammonia 생성능을 부가함으로써 생물방제력의 상승효과를 거둘 수 있었다.

• 한국식품영양학회지
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• 제21권1호
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• pp.15-21
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• 2008

In this study, the minimum inhibition concentration(MIC), growth inhibition activity, and colony forming inhibitory activity of water soluble propolis against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae and Salmonella enteritidis were tested. The MICs of the water soluble propolis against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, and Salmonella enteritidis were 312.5 ppm, below 156.3 ppm, 625 ppm, 10,000 ppm, above 10,000 ppm, 10,000 ppm, above 10,000 ppm, above 10,000 ppm, 10,000 ppm, and above 10,000 ppm, respectively. The growth inhibition concentrations against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, and Klebsiella pneumoniae were 156.3 ppm, below 156.3 ppm, 625 ppm, 5,000 ppm, 10,000 ppm, 10,000 ppm, 10,000 ppm, 10,000 ppm, and 5,000 ppm, respectively. However, 10,000 ppm did not inhibit the growth of Salmonella enteritidis. Finally, the colony forming inhibitory activities against Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Streptococcus mutans, Citrobacter freundii, Enterobacter aerogenes, Escherichia coli, Klebsiella pneumoniae, and Salmonella enteritidis were 98.0%, 99.8%, 69.8%, 98.1%, 62.0%, 63.1%, 79.5%, 61.9%, 79.6%, and 0.0%, respectively.

• KSBB Journal
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• 제13권5호
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• pp.491-496
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• 1998

The production of fibrinolytic enzyme from Bacillus subtilis BK-17 was studied in the shake flask cultures. The important medium components studied include nitrogen source, carbon source and inorganic salts. The environmental conditions include initial pH, temperature, shaking speed and working volume. Among various N-sources, C-sources and inorganic salts tested, soybean flour, D-glucose and Na2HPO4 gave the best results, and their optimal concentrations were 1.5%, 0.5% and 0.05%, respectively. The optimal pH and temperature were 9.0 and 37$^{\circ}C$. With decreasing working volume in the range of 25∼100ml in the 250ml flask or increasing shaking speed in the range of 100∼300rpm, the enzyme production was greatly enhanced. The enzyme activity under the optimal conditions was about 1400I.U./ml with urokinase as a standard.