• Title, Summary, Keyword: B. subtilis

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High-Level Expression and Secretion of Bacillus pumilus Lipase B26 in Bacillus subtilis Chungkookjang

  • Lee, Mi-Hwa;Song, Jae-Jun;Choi, Yoon-Ho;Hong, Seung-Pyo;Rha, Eu-Gene;Kim, Hyung-Kwoun;Lee, Seung-Goo;Poo, Har-Young;Lee, Sang-Chul;Seu, Young-Bae;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.892-896
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    • 2003
  • High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

Characteristics of Cadmium-Resistant Bacillus subtilis DT134 (Bacillus subtilis DT134의 카드뮴 저항성)

  • 윤경표
    • KSBB Journal
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    • v.13 no.4
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    • pp.383-390
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    • 1998
  • Bacillus subtilis DT134 was resistant to 50-fold higher concentration of cadmium ions (Cd2+) than cadmium-sensitive B. subtilis BD224 in Luria Broth (LB) medium. Minimal inhibition concentration test in LB agar plates also showed similar results. The elevated cadmium resistance of B. subtilis DT134 strongly suggested a possible existence of cadmium resistance gene in it. Southern blot with Staphylococcus aureus cadA gene fragment (757 bp NlaIV-XmnI cadA DNA fragment) as probe was carried out to test the existence and similarity of the gene. In high stringency condition, there was no detectable signal, but in low stringency, a strong signal specific to the cadA probe could be detected. These results strongly suggested that there was some similarity between total DNA of B. subtilis DT134 and S. aureus pl258 in terms of cadmium resistance gene and the resistance mechanism might be an efflux mechanism. The subsequent efflux experiment showed that the cadmium resistance mechanism of B. subtilis DT134 was also due to the efflux of cadmium.

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Biological Activity and Biochemical Properties of Silkworm (Bombyx mori L.) Powder Fermented with Bacillus subtilis and Aspergillus kawachii (유용식용 균주에 의한 발효 누에분말의 이화학적 특성과 생리활성)

  • Cha, Jae-Young;Kim, Yong-Soon;Ahn, Hee-Young;Kang, Min-Jung;Heo, Su-Jin;Cho, Young-Su
    • Journal of Life Science
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    • v.21 no.1
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    • pp.81-88
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    • 2011
  • Biological activities (${\alpha},{\alpha}'$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) free radical scavenging activity, fibrinolytic activity and reducing power) and biochemical properties (protein content and electrophoretical protein patterns) were examined in solid state fermentation with Bacillus subtilis and Aspergillus kawachii using silkworm powder (SP) as substrate. The highest protein contents and free radical scavenging activities were seen in the SP fermented for 12 days with B. subtilis and A. kawachii, and these were in a time-dependent manner. The highest reducing power was seen in the SP fermented for 6 days with B. subtilis and for 12 days with A. kawachii, respectively. The highest fibrinolytic activities were seen in silkworm fermented for 6 days with B. subtilis and A. kawachii, but this activity was higher in the A. kawachii fermented SP than that of B. subtilis. When total protein patterns were analyzed by SDS-polyacrylamide gel electrophoresis (PAGE), the proteins of the SP fermented with B. subtilis for 3 days were completely degraded, while the protein degradation in the SP fermented with A. kawachii occurred after 12 days and this degradation increased proportionally to culture time. As a result, the SP fermented with both B. subtilis and A. kawachii showed higher fibrinolytic activities after 6 days of fermentation and antioxidative activity after 12 days, indicating that physiological activities of the fermented SP using these strains were highly improved compared to the unfermented SP, and that this compound could be a candidate material as a dietary supplement of healthy functional foods.

Functional Analysis of Bacillus subtilis Isolates and Biological Control of Red Pepper Powdery Mildew Using Bacillus subtilis R2-1 (Bacillus subtilis 균주의 기능성 분석 및 이를 활용한 고추 흰가루병 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Choi, Eun-Jung;Lee, Min-Ho;Park, Jong-Ho;Han, Eun-Jung;An, Nan-Hee;Jee, Hyeong-Jin
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.201-209
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    • 2012
  • The multi-function of 18 Bacillus subtilis isolates collected from agricultural extension centers of local government and National Academy of Agricultural Science was investigated by measuring their antifungal activities against five plant pathogens, such as Rhizoctonia solani, Colletotrichum acutatum, Fusarium oxysporum, Magnaporthe oryzae and Phytophthora capsici, phosphorus solubilization ability, production of indole acetic acid (IAA) and siderophore, and nitrogen fixation. The B. subtilis isolates showed antifungal activity against several plant pathogens and nitrogen fixation activity, and produced siderophore and IAA. They could control pepper powdery mildew (Leveillula taurica), but there was no difference in control efficacy among the B. subtilis isolates. In fields, the control efficacy of B. subtilis R2-1 ($10^8$ cells/ml) was compared with two microbial fungicides, Q-pect and Topsid. In 2009, the control efficacy of B. subtilis R2-1 (37.7%) was lower than that of Topsid (47.6%), but higher than that of Q-pect (25.7%). In 2010, the control efficacy of B. subtilis R2-1 (83.3%) was higher than that of Topsid (67.9%). In order to elucidate mode of action of B. subtilis R2-1 for controlling pepper powdery mildew, spore germination rates of pepper powdery mildew pathogen collected on treated leaves was investigated when suspensions of B. subtilis R2-1 and two microbial fungicides (Q-pect and Topsid) were foliar-sprayed. They highly suppressed spore germination of the pathogen with inhibition values of 84.2% for B. subtilis R2-1, 97.9% for Q-pect and 94.7% for Topsid. Further study on the mass-culturing method and formulation is needed for development of a microbial fungicide.

The Antioxidant Effect of Cheonggukjang, Fermented Using the New Strain, Bacillus amyloliquefaciens NBF11-1 (신 균주 Bacillus amyloliquefaciens NBF11-1을 이용하여 발효한 청국장의 항산화효과)

  • Kim, Han Soo;Yun, Hyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.8
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    • pp.5343-5350
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    • 2015
  • This study aims to compare and analyze the antioxidant effect of Cheonggukjang's traditional fermentation strain, Bacillus subtilis NG24 which was a control of the study, in order to see the biological activity effect of the new strain, Bacillus amyloliquefaciens NBF11-1 that was first found in the surface of the bamboo stem, but hasn't been insufficiently researched. In the antioxidant activity experiment of the Cheonggukjang extract, the B.amyloliquefaciens NBF11-1 sample showed a significant increase in the total polyphenol extract content, compared to B.subtilis NG24(p=0.032). Also, compared to B.subtilis NG24, the sample containing B.amyloliquefaciens NBF11-1 showed a significant increase in SOD-like Activity, DPPH radical scavenging, and NO radical scavenging, as the concentration rose(p<.05). Additionally, $IC_{50}$ in each antioxidant activity experiment significantly decreased in the B.amyloliquefaciens NBF11-1 sample like SOD-like Activity(p=0.045), DPPH radical scavenging(p=0.041), and NO radical scavenging(p=0.019), compared to B.subtilis NG24.

Functional Mechanism of Plant Growth Retardation by Bacillus subtilis IJ-31 and Its Allelochemicals

  • Kim, Won-Chan;Rhee, In-Koo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1375-1380
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    • 2012
  • We previously isolated a rhizobacterium (Bacillus subtilis IJ-31) and demonstrated that its associated allelochemicals could indicate plant growth retardation. However, little is known about how the growth of plants is regulated by B. subtilis IJ-31 and its allelochemicals. In this study, we investigated whether plant growth retardation in this relationship occurred through the inhibition of gibberellin (GA) biosynthesis. GA $3{\beta}$-hydroxylase activity was found to be inhibited by B. subtilis IJ-31 and hydrocinnamic acid (HCA), which is one of the allelochemicals produced by B. subtilis IJ-31. Additionally, thin layer chromatography (TLC) demonstrated that B. subtilis IJ-31 culture broth and HCA both inhibit GA $3{\beta}$-hydroxylase (MBP-GA4) activity. The retardation of plants by HCA was then confirmed in vivo and in vitro using a Ryegrass and Arabidopsis growth retardation assay. Furthermore, treatment with either B. subtilis IJ-31 culture extract or its allelochemicals resulted in significant down-regulation of XTR9 gene expression in Arabidopsis. Overall, we identified the functional mechanism of plant growth retardation by B. subtilis IJ-31 and its allelochemicals.

Application of Bacillus subtilis 168 as a Multifunctional Agent for Improvement of the Durability of Cement Mortar

  • Park, Sung-Jin;Park, Jong-Myong;Kim, Wha-Jung;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1568-1574
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    • 2012
  • Microbiological calcium carbonate precipitation (MCCP) has been investigated for its ability to improve the durability of cement mortar. However, very few strains have been applied to crack remediation and strengthening of cementitious materials. In this study, we report the biodeposition of Bacillus subtilis 168 and its ability to enhance the durability of cement material. B. subtilis 168 was applied to the surface of cement specimens. The results showed a new layer of deposited organic-inorganic composites on the surface of the cement paste. In addition, the water permeability of the cement paste treated with B. subtilis 168 was lower than that of non-treated specimens. Furthermore, artificial cracks in the cement paste were completely remediated by the biodeposition of B. subtilis 168. The compressive strength of cement mortar treated with B. subtilis 168 increased by about 19.5% when compared with samples completed with only B4 medium. Taken together, these findings suggest that the biodeposition of B. subtilis 168 could be used as a sealing and coating agent to improve the strength and water resistance of concrete. This is the first paper to report the application of Bacillus subtilis 168 for its ability to improve the durability of cement mortar through calcium carbonate precipitation.

Effect of Bacillus subtilis Natto on Meat Quality and Skatole Content in TOPIGS Pigs

  • Sheng, Q.K.;Zhou, K.F.;Hu, H.M.;Zhao, H.B.;Zhang, Y.;Ying, W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.5
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    • pp.716-721
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    • 2016
  • This study investigated the effect of Bacillus subtilis (B. subtilis) natto on meat quality and skatole in TOPIGS pigs. Sixty TOPIGS pigs were randomly assigned to 3 groups (including 5 pens per group, with 4 pigs in each pen) and fed with basic diet (control group), basic diet plus 0.1% B. subtilis natto (B group), and basic diet plus 0.1% B. subtilis natto plus 0.1% B. coagulans (BB group), respectively. All pigs were sacrificed at 100 kg. Growth performance, meat quality, serum parameters and oxidation status in the three groups were assessed and compared. Most parameters regarding growth performance and meat quality were not significantly different among the three groups. However, compared with the control group, meat $pH_{24}$, fat and feces skatole and the content of Escherichia coli (E. Coli), Clostridium, $NH_3$-N were significantly reduced in the B and BB groups, while serum total cholesterol, high density lipoprotein, the levels of liver P450, CYP2A6, and CYP2E1, total antioxidant capability (T-AOC) and glutathione peroxidase and Lactobacilli in feces were significantly increased in the B and BB groups. Further, the combined supplementation of B. subtilis natto and B. coagulans showed more significant effects on the parameters above compared with B. subtilis, and Clostridium, and $NH_3$-N. Our results indicate that the supplementation of pig feed with B. subtilis natto significantly improves meat quality and flavor, while its combination with B. coagulans enhanced these effects.

Evaluation of Disinfection Characteristics of Ozone, UV Processes for Bacillus Subtilis Spores Inactivation (Bacillus Subtilis Spores 불활성화 실험을 통한 오존, UV 공정의 소독 특성 평가)

  • Jung, Yeon Jung;Oh, Byung Soo;Kang, Joon-Wun
    • Journal of Korean Society on Water Environment
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    • v.22 no.4
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    • pp.672-677
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    • 2006
  • Ozone/UV combined process is an effective technique to enhance generation of OH radical which is non-selective and powerful oxidant. The objective of this study is to evaluate the inactivation rates of B. subtilis spores by three candidate processes (ozone alone, UV alone, ozone/UV combined processes) at 4 and $20^{\circ}$ and to investigate the effects of OH radical on inactivation of B. subtilis spores. On the UV alone process, required UV dosages for lag phase and 3-log inactivation of B. subtilis spores were determined as $8.9mJ/cm^2$ and $47mJ/cm^2$. However, the inactivation of B. subtilis spores didn't occured beyond 4.5-log inactivation despite increasing UV dose. The inactivation of B. subtilis spores by ozone alone and ozone/UV combined process was investigated with ozone CT (Concentration of disinfectant ${\times}$ Contact time) concept. As a result, inactivation of B. subtilis spores by ozone/UV combined process was faster than by ozone alone, and especially $CT_{lag}$ value B. subtilis spores in the presence and absence of t-BuOH, OH radical scavenger, was investigated to evaluate effects of OH radical formed during ozone/UV combined process. We found that OH radical plays important roles on inactivation of B. subtilis spores.

Characterization of ${\gamma}$-Polyglutamic Acid Produced from the Solid-state Fermentation of Soybean Milk Cake Using Bacillus sp.

  • Oh, Soo-Myung;Jang, Eun-Kyung;Seo, Ji-Hyun;Ryu, Mi-Jin;Lee, Sam-Pin
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.509-514
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    • 2007
  • In this study, we optimized the production of ${\gamma}-polyglutamic$ acid (PGA) in soybean milk cakes (SMC) fermented with Bacillus subtilis GT-D and B. subtilis KU-A, to be utilized as a functional food ingredient. PGA production was dependent upon the glutamate content, fermentation time, and type of Bacillus sp. The consistencies of the SMCs fermented by B. subtilis GT-D and B. subtilis KU-A were highest after 36 hr of fermentation, and then decreased gradually. The SMC fermented by B. subtilis KU-A had a higher consistency than the SMC fermented by B. subtilis GT-D. In the presence of 10% defatted soy flour (DFS), 5% glutamate in the SMC was efficiently converted into polyglutamic acid (PGA) for 24 hr, indicating a conversion yield above 96%, but its conversion then decreased with higher concentrations of glutamate. The soluble solid content (mucilage) of the SMC fermented with B. subtilis KU-A was 9.5%(w/w), and composed of 65.6% PGA (Mw 1,536 kDa) and some polysaccharides. However, the SMC fermented with B. subtilis GT-D had a mucilage content of 7.8%(w/w), and was composed of 66.4% PGA (Mw 1,409 kDa), 11.5% levan, and some polysaccharides. The viscoelastic values of the mucilage obtained using B. subtilis KU-A were much higher than those of mucilage obtained using B. subtilis GT-D. Also, the G'-value (elastic modulus) was higher than the G"-value (viscous modulus).