• 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.

• 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).

• Journal of Applied Biological Chemistry
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• v.52 no.3
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• pp.121-125
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• 2009

We measured the influence of antifungal antagonists Bacillus subtilis AH18 and Bacillus licheniformis K11 on soil microbial community in microcosms. Both antifungal antagonists were confirmed to suppress hot pepper phytophthora blight. Phospholipid fatty acids (PLFA) were analyzed to investigate the soil microbial community. B. subtilis AH18 changed the total PLFA composition and bio-indicators of PLFA, compared with other treatments. B. subtilis AH18 decreased the proportion of bacteria and gram negative/gram positive bacteria, and increased the fungi/bacteria and anaerobic/aerobic microorganisms. In addition cy19:0/18:$1{\omega}7c$, which means adaptation to unfavorable environmental conditions, was increased by the application of B. subtilis AH18. On the other hand the inoculation of B. licheniformis K11 or combined inoculation of both antifungal strains did not affect soil microbial community. The suppression of phytophthora blight and preservation of indigenous soil microbial community may be achieved by the combined application of B. subtilis AH18 and B. licheniformis K11.

• Korean Journal of Food Science and Technology
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• v.41 no.4
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• pp.399-404
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• 2009

The goal of this study was to improve the quality of cheonggukjang by the optimization of the inoculation methods of the Bacillus subtilis (B. subtilis) and Lactobacillus plantarum (L. plantarum) strains. In order to optimize the mixed cultivation of B. subtilis and L. plantarum, the B. subtilis strain was inoculated into steamed soybeans after cultivation of L. plantarum. Inoculation size of B. subtilis was changed to the simultaneous inoculation method in order to stimulate the growth of the L. plantarum in cheonggukjang. The viable cell count of L. plantarum increased from $2{\times}10^7$ CFU/g to $2-6{\times}10^8$ CFU/g and B. subtilis grew to $9{\times}10^8$ CFU/g. These results showed that 2 strains were successfully able to grow in the steamed soybean for good quality of cheonggukjang by optimization of the inoculation methods. The sensory evaluation indicated that a favorable aroma and overall acceptance of cheonggukjang by the optimized mixed cultivation of B. subtilis and L. plantarum, which was relatively higher than those of cheonggukjang by single strain inoculation of B. subtilis.

• Journal of Animal Science and Technology
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• v.52 no.5
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• pp.389-398
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• 2010

The objective of this study was to investigate the effects of three strains of Bacillus subtilis (B. subtilis) supplemented to diets on egg production, egg quality, egg yolk cholesterol levels, the profile of cecal microflora, and tibia characteristics in laying hens. One hundred sixty 76-week-old Hy-Line Brown layers were randomly divided into 4 groups with 4 replicates per group (10 birds per replicate). Birds in the control group were fed a corn-soybean meal based diet. The remaining three treated groups were fed the control diet containing either 0.05% B. subtilis Ch3 (T1), 0.05% B. subtilis Ch3 + B. subtilis W1 (T2) or 0.05% B. subtilis commercial product (T3) for 6 weeks, respectively. There were no differences in feed intake, egg weight, egg production and egg mass among the groups. The dietary supplementation of B. subtilis improved eggshell strength and Haugh units compared to those of control (P<0.05). The activities of GOT and GPT in serum were not also affected by the dietary treatments. The population of total microbes and lactic acid bacteria in cecum were significantly increased by the dietary B. subtilis (P<0.05), but not the coliforms. The cholesterol concentration in egg yolk and serum in the treated groups were significantly decreased compared to those of control (P<0.05). Also, The levels of phospholipids in serum were significantly decreased compared to those of control (P<0.05). The supplementation of three strains of B. subtilis to diets significantly increased the contents of tibia ash compared to that of control (P<0.05). Thus, this study showed significant improvements in egg quality, such as eggshell strength and Haugh unit, by dietary B. subtilis strains. The B. subtilis strains added to the diets modulated the profiles of cecal microflora, reflecting beneficial effects in laying hens.

• The Korean Journal of Community Living Science
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• v.18 no.3
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• pp.399-406
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• 2007

Soybean seeds were fermented by Bacillus subtilis and/or Lactobacillus bulgaricus to improve solubility, viscosity, water holding capacity and oil holding capacity of soybean proteins in Chongkukjang. The maximum colony forming unit and protease activity of B. subtilis or L bulgaricus were observed after 60 hours of fermentation, and those of the mixed fermentation by two microorganisms were steadily increased during the fermentation periods. Solubilities of soybean proteins by B. subtilis or L bulgaricus were steadily increased before the values were considerably increased to 60 hours of fermentation, whereas water holding capacities of the proteins were decreased by B. subtilis or L. bulgaricus and those of the mixed fermentation were decreased progressively. Viscosities of soybean proteins by B. subtilis and/or L. bulgaricus were decreased progressively during the fermentation. Viscosities of soybean proteins by B. subtilis and/or L. bulgaricus were decreased progressively during the fermentation. Oil holding capacities of soybeans by B. subtilis or L. bulgaricus were maximum at 20 or 80 hours of fermentation and those of the mixed fermentation were decreased after 10 hours of the fermentation.

• Korean Journal of Veterinary Service
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• v.29 no.3
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• pp.257-266
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• 2006

The present study was done to investigate the effect of dietary supplemental freezer dry powder fermentation soybean by Bacillus subtilis (natto) on the growth performance and intestinal microflora, prevention of fowl typhoid infection, the uptake of vegetative diet in broiler chickens. The chickens were fed control diet (supplement antibiotics) and fermentation soybean (0.75, 1.5, 3%) diets. A total of 280 one day old broiler chickens with randomly mixed sexes were fed the four diets for 6 weeks. Body weight gain of chicken fed 1.5% fermentation soybean by B subtilis tend to increase higher than the control from 6th week. Chickens fed diets containing 1.5% fermentation soybean by B subtilis had higher intake than those fed the other levels to the 3th weeks, but lower than control from the 4th week to the 6th week. Feed conversion also improved significantly in the supplemental 1.5% fermentation soybean by B subtilis from the 4th week to the 6th week. The number of B subtilis and Lactobacillus spp in the ileum and cecum tend to increase in the supplemental fermentation soybean by B subtilis at 6 week of age, but was not significantly different. In the nutrient digestibility, the feed conversion on the supplemental 1.5% fermentation soybean by B subtilis was better than the control and the weight of drying feces lower than the control. In test of S. gallinarum intramuscular inoculation, reisolation rate of S gallinarum in liver and feces 1.5% the fermentation soybean by B subtilis 75% (liver), 17% (feces) had decreased than the control.

• Journal of the East Asian Society of Dietary Life
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• v.14 no.4
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• pp.363-369
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• 2004

Three different doengjang(fermented soybean paste) were prepared by using B. subtilis SS103 with high angiotensin converting enzyme(ACE) inhibitory activity, Asp. oryzae and mixed culture of two organisms, respectively. Quality characteristics and the inhibitory activity were compared. $\beta$-Amylase activity rapidly increased during 30 days of fermentation, but lower activity was found in doenjang prepared with B. subtilis. However, $\alpha$-Galactosidase activity decreased for 15 days of fermentation and little activity was observed after that fermentation period. Protease production tended to increase during the fermentation. High protease activity was found only in doengjang prepared with Asp. oryzae, but the doengjang prepared with B. subtilis SS103 showed very low activity. Total isoflavone content at 90 day fermentation was the highest in doengjang prepared with mixture of Asp. oryzae and B. subtilis. ACE inhibitory activity increased during the fermentation period. The highest activity was found in doengjang made with mixture of Asp. oryzae and B. subtilis. Sensory evaluation on doengjang itself and its soup indicated that doengjang made with mixture of Asp. oryzae and B. subtilis showed higher acceptability on taste, flavor and color than those prepared with only Asp. oryzae or B. subtilis, respectively.

• Journal of the Korean Applied Science and Technology
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• v.37 no.6
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• pp.1535-1544
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• 2020

The primary objective of this study was to isolate and identify amine oxidase-producing probiotic Bacillus strains from traditional fermented soybean paste. Biogenic amines (BA)-forming bacteria isolated from the samples were identified as Bacillus sp. TS09, Bacillus licheniformis TS17, Bacillus subtilis TS19, Bacillus cereus TS23, Bacillus sp. TS30, Bacillus megaterium TS31, B. subtilis TS44, Bacillus coagulans TS46 and Bacillus amyloliquefaciens TS59. Meanwhile, B. subtilis TS04 and TS50 isolated from the same samples exhibited good probiotic properties, including the tolerance to artificial gastric juice and bile salts, the adhesion to intestinal epithelial cells, and the production of bacteriocin(s) active against BA-forming bacteria (Bacillus sp. TS30 and B. subtilis TS44). In addition, the amine oxidase produced by B. subtilis TS04 and TS50 significantly decreased the formation of BA, especially cadaverine, putrescine, and tyramine, therefore, these strains could be considered good potential probiotic candidates to prevent or reduce BA accumulation in food products.