• 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.

• 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.

• 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.

• 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.

• 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.

• 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.

• Food Science and Preservation
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• v.20 no.3
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• pp.379-385
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• 2013

To establish the optimal manufacturing conditions of soybean koji, soybean Koji prepared with Aspergillus oryzae 6-M-1 and Bacillus subtilis 3-B-1 isolated from traditional Korean meju. During 7 days of making Koji, the amount of amino-type nitrogen was getting more increase. The amount of amino-type nitrogen of Koji prepared with A. oryzae 6-M-1 was 686.16 mg% (w/w), that of Koji with B. subtilis 3-B-1 was 643.46 mg% (w/w) at seventh day of making Koji. The ${\alpha}$-amylase activity of Koji prepared with A. oryzae 6-M-1 was 1472.54 unit/g, that of Koji with B. subtilis 3-B-1 was 791.00 units/g on the seventh day of the making. The acidic protease activity of Koji prepared with A. oryzae 6-M-1 was 309.00 unit/g, that of Koji with B. subtilis 3-B-1 was 135.88 unit/g at 7th day of making. The amount of amino-type nitrogen and enzyme activities of soybean Koji prepared with A. oryzae 6-M-1 and B. subtilis 3-B-1 were produced more than those of wheat flour Koji made in factory. Sensory evaluation on a commercial doenjang and doenjangs prepared with A. oryzae 6-M-1 and B. subtilis 3-B-1 was not significantly different at p<0.05.

• 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.

• Journal of Microbiology and Biotechnology
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• v.32 no.8
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• pp.1011-1016
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• 2022

Bacillus subtilis is a useful bacterium in the food industry with applications as a starter strain for fermented food and as a probiotic. However, it is difficult to discriminate B. subtilis from other Bacillus species because of high phenotypic and genetic similarity. In this study, we employed five previously constructed multilocus sequence typing (MLST) methods for the discrimination of B. subtilis from other Bacillus species and all five MLST assays clearly distinguished B. subtilis. Additionally, the 17 housekeeping genes used in the five MLST assays also clearly distinguished B. subtilis. The pyruvate carboxylase (pyrA) and shikimate dehydrogenase (aroE) genes were selected for the discrimination of B. subtilis because of their high number of polymorphic sites and the fact that they displayed the lowest homology among the 17 housekeeping genes. Specific primer sets for the pyrA and aroE genes were designed and PCR products were specifically amplified from B. subtilis, demonstrating the high specificity of the two housekeeping genes for B. subtilis. This species-specific PCR method provides a quick, simple, powerful, and reliable alternative to conventional methods in the detection and identification of B. subtilis.

• 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.