• 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.

• 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.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.522-529
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• 2019

A Bacillus strain, SJ4, exhibiting strong fibrinolytic activity was isolated from saeu (shrimp, Acetes chinensis) jeotgal, a Korean traditional fermented food and was identified as B. subtilis. The B. subtilis SJ4 strain can grow at a NaCl concentration of up to 15% (w/v). The fibrinolytic activity of B. subtilis SJ4 (152.0 U/ml) cultured in Luria-Bertani (LB) broth for 48 h at 37℃ with aeration was higher than that of B. subtilis SJ4 cultured in TSB (124.5 U/ml) under same culture conditions. The major proteins in the LB culture supernatant of B. subtilis SJ4 were analyzed by SDS-PAGE, which revealed three major bands (23, 25, and 28 kDa). The band (23 kDa) with strong fibrinolytic activity, analyzed on fibrin zymogram, was observed at 60-96 h of cultivation. The aprESJ4 gene encoding the major fibrinolytic enzyme, AprESJ4, was cloned by PCR. The aprESJ4 gene sequence exhibited high similarities with the fibrinolytic gene sequences of other Bacillus species. The amino acid sequence of AprESJ4 exhibited 98.9 and 98.4% similarity with subtilisin NAT and AprE2 of B. subtilis, respectively. Hence, B. subtilis SJ4 can be a potential starter culture for jeotgal products.

• 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.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.50-55
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• 2001

A new bacteriocin produced by Bacillus subtilis cx1, was partially purified and characterized. The bactericoin from B. subtilis cx1 was stable in the range of pH 2.5-9.5. B. subtilis csx1 retained its antimicrobial activity to long-term exposure at $-20^{\circ}C$ and $-70^{\circ}C$. However, B. subtilis cx1 was inactivated completely within 15 min over $60^{\circ}C$ and lost 50% of its antimicrobial activity within 15 min at $50^{\circ}C$, B. subtilis cx1 was inactivated by protease, trypsin, proteinase K and carboxypeptidase, which indi-cates its protein nature. Direct detection of the antimicrobial activity on Tricine -SDS-PAGE suggested an apparent molecular mass of about 9,500 dalton.

• Food Science and Preservation
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• v.24 no.7
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• pp.975-982
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• 2017

Health of human intestine has deteriorated due to excessive stress and western diet. In this study, Vigna angularis was fermented by Bacillus subtilis KCCM11965P in order to develop prebiotic resource for improving bowel movement. The contents of ash and crude protein were $3.35{\pm}0.04%$ and $21.1{\pm}0.19%$ respectively. Vigna angularis extract (1, 3, 5%) were incubated with 3% (v/v) Bacillus subtilis KCCM 11965P for 0, 24, 48, and 72 h. Total bacterial numbers showed that the combination of 3% powdered seeds and 72 h incubation time was optimum condition for this experiment. Total polyphenol content increased from $0.18{\pm}0.010mg/mL$ in pre-incubation to $0.23{\pm}0.007mg/mL$ in post-incubation with the condition mentioned above. DPPH radical scavenging activity also increased from $36.1{\pm}6.0%$ to $63.6{\pm}5.2%$. Analysis of protease activity showed $2.69{\pm}0.003unit/mL$ in combination of 5% powdered seeds and 72 h incubation time. Amylase activity increased from $1.0{\pm}0.1unit/mL$ in pre-incubation to $26.0{\pm}0.2unit/mL$ in post-incubation. The analysis of free amino acids after incubation with Bacillus subtilis KCCM 11965P showed that leucine increased from 5.22 mg/L to 67.59 mg/L and tyrosine, one of non-essential amino acid also increased 10.08 mg/L to 259.35 mg/L by incubation with 5% powdered seeds. Most of organic acid were reduced by incubation for 72 h. These results suggest that Vigna angularis could be utilized most as a prebiotic resources.

• Food Science and Biotechnology
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• v.18 no.4
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• pp.1035-1037
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• 2009

In this study we analyzed the dynamic changes in microbiota composition during kochujang fermentation at $30^{\circ}C$. During fermentation, the viable cell counts slowly increased and reached $3.2{\times}10^7$ for aerobic bacteria, $8.3{\times}10^3$ for yeast, and $1.4{\times}10^3$ CFU/mL for fungi after 60 days. Bacilli were found to be the most dominant microorganisms throughout the fermentation process. Using the culture dependent method Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloquefaciens were found to be the main species during the early stages of fermentation; however, Bacillus pumilus and Bacillus stearothermophilus became the most dominant species during the late stage of fermentation. In contrast, when the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was used Bacillus ehimensis was found to be the dominant species during the early stage of fermentation and Bacillus megaterium, B. pumilus, B. subtilis, and B. licheniformis were dominant in the ate stages. These results indicate various other Bacillus species rather than just B. subtilis and B. licheniformis might be involved in the fermentation of kochujang.

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.655-659
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• 1998

Bacterial strain showing the strong fibrinolytic activity (2.04 plasmin unit) was screened from Jeot-Gal, Korean salt-fermented fish collected from various region. For the identification, when the strain was characterized morphologically, culturally, and biochemically, it was identified to Bacillus pumilus. And, when the fatty acids composition of the strain was analyzed, it was identified to Bacillus atropheus. Finally, the 16S rRNA partial sequence (V3 region) showed that the fibrinolytic stain screened from Jeot-Gal was identified as Bacillus subtilis. So, we named it Bacillus subtilis KJ-48.

• Korean Journal of Microbiology
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• v.36 no.3
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• pp.203-208
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• 2000

A Baczllus circdans KCTC3004 $\alpha$-amylase gene contained in a recombinant plasmid pAL850 was transferred into a new shuttle vector plasmid pALSIlI by ligating linearlzed DNAs of pUC19 and pUB110. B. subtilis RM125 and B. megatenurn ATCC14945 transfonned with pALS111 produced the $\alpha$-amylase substantially Most of the enzyme was produced during the exponential growth period. The maxiinurn activities of the $\alpha$-amylase produced by the Bucillus transformants were compared with that of the B. circulans gene donor strain. The B. subtilis RM125(pALS111) enzyme showed the actlvicy 95 times higher than that of the gene donor cells, followed by the B, nzegaterium ATCC14945(pALSlll) enzyme with activity 34 limes higher than that of the gene donor cells. While E coli secreted about 10% of the produced enzyme, B. subtilis excreted the enzyme inlo the medium wholly and B. megaterirun about 98% ofthe total product. The plasmid pALSI11 was quite stable inB. nzegaterium (92%), inoderately stable in B. subtilis (76%), but was unstable in E. coli (38%). The SDS-PAGE and zymogram of this enzyme produced in E. coli(pALS111), B. subtilis( pALS111) or B. megateril~m (pALS111) indicated a molecular weight of 55,000. The enzymes overproduced in three different host cells hydrolyzed starch to produce mainly maltoaiose and mallooligosaccharides.

• Korean Journal of Organic Agriculture
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• v.19 no.3
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• pp.385-396
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• 2011

This study was carried out to develop environmental friendly control for major diseases and pests on Boxthorn (Lycium chinense Mill.). Outbreak of Eighteen diseases and pests were found at the Boxthorn organic yards in Chung-nam province. Among them Powdery mildew (Erysiphe polygoni de Cand.), Hypophyllous mold (Pseudocercospora chengtuensis (Tai)), Western flower trips (Frankliniella occidentalis (Pergande)), Green peach aphid (Myzus pericae (Sulzer)) and Corn earworm (Helicoverpa armigera) needed to be controled by environmental friendly methods for high fruit yield of organic Boxthorn. In summer(Jun) test Bacilus subtilis QST 713 wettable powder and Sulfur wettable powder were effective and in autumn (Sep.) test Sulfur, Copper hydroxide and Paraffinic oil were relatively effective in Powdery mildew. In Hypophyllous mold control test Paraffinic oil and Bacilus subtilis GB - 0365 were effective with above 70% control value. And it was possible to control Western flower trips by natural enemy (Orius laevigatus) by 80% control value. Corn earworm was possible to control by Bacillus thuringiensis subsp. aizawai GB413 flowable and Bacillus thuringiensis aizawa 0423 wettable powder application above 70%. And Green peach aphid was controllable with environmental friendly materials, such as, Bacillus subtilis (Seoncho), Bacillus subtilis (Jinsami) above 80% and Ginkgo nut extract above 70% control value.