• Korean Journal of Food Science and Technology
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• v.44 no.3
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• pp.312-316
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• 2012

In order to understand the effect of hydrostatic pressure (HP) on Bacillus subtilis isolated from makgeolli, the survival of B. subtilis after HP treatment (400 MPa for 5 min) in various substrates including phosphate buffer, tryptone soya broth at pH 7 and 4, and makgeolli at pH 4 was evaluated depending on bacterial forms (spores and vegetative cells) and adaptation conditions ($25^{\circ}C$ for 3 h, or $10^{\circ}C$ for 24 h). Spores were generally resistant to HP (<1 log reduction) regardless of conditions. In contrast, vegetative cells were generally susceptible to HP (up to 3 log reduction-except makgeolli) and were more susceptible after 3 h at $25^{\circ}C$ compared to 24 h at $10^{\circ}C$. In vegetative cells inoculated makgeolli (7 log CFU/mL), the colonies were not detected after 24 h at $10^{\circ}C$. Consequently, B. subtilis in makgeolli easily existed as spores and the spores were resistant to HP. Results demonstrate that HP was more promising in the inactivation of vegetative cells.

• Food Science of Animal Resources
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• v.28 no.2
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• pp.181-186
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• 2008

This study was conducted to investigate the effects of supplementation with transgenic Bacillus subtilis secreting chitinase on the growth performance, nutrient digestibility, blood characteristics and carcass traits in finishing pigs. A total of sixty-four pigs ($50.82{\pm}0.82kg$, average initial body weight) were assessed over a period of 84 days. Dietary treatments included: 1) CON (basal diet without antibiotics), 2) AD (basal diet + 0.1% Virginiamycin), 3) CD0.5 (basal diet + 0.5% transgenic B. subtilis), 4) CD1.0 (basal diet + 1.0% transgenic B. subtilis). Each dietary treatment had 4 replicates of 4 pigs per pen in a randomized complete block design. In terms of growth performance, the ADG (average daily gain) and gain/feed ratio were significantly increased with the CD1.0 diet compared to the AD diet during weeks 0-4 (p<0.05). During weeks 4-8, the ADG was significantly increased with the AD diet compared to the CON and CD0.5 diets (p<0.05). The ADFI was also significantly increased with the AD diet compared to the other diets (p<0.05). During weeks $8{\sim}12$, the ADFI was significantly increased with the CON diet compared to the other diets (p<0.05), and the gain/feed ratio was significantly increased with the CD0.5 and CD1.0 diets compared to the AD diet (p<0.05). Over the entire 84 day test period, the ADFI was significantly increased with the AD diet compared to the CD0.5 and CD1.0 diets (p<0.05). The gain/feed ratio was significantly increased with the CD0.5 and CD1.0 diets compared to the CON diet (p<0.05). In terms of meat color, the L value was significantly increased with the CD0.5 diet compared to the CON and AD diets (p<0.05), and the a value was significantly increased with the CON diet compared to the other diets (p<0.05). In terms of sensory evaluation, meat color was significantly improved with the CON, CD0.5 and CD1.0 diets compared to the AD diet (p<0.05). Marbling was significantly increased with the CON diet compared to the other diets (p<0.05). Firmness was significantly increased with the CD0.5 diet compared to the AD diet (p<0.05). In conclusion, supplementation with transformed B. subtilis secreting chitinase improved gain/feed ratios and influenced meat color. Thus, we suggest that transformed B. subtilis secreting chitinase can partially substitute for antibiotics.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.498-508
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• 2019

Bacillus species were isolated from iru, a traditional fermented condiment in Nigeria. Polyphasic approach was used to evaluate the phylogenetic relationship and strain sub-type of the isolated species. Additionally, the phylogenetic profiles of the species isolated from iru were compared with those of bacilli isolated from different continents. The phylogenetic diversity analysis was performed using the combination of 16S rRNA gene sequencing, ITS-PCR, ITS-PCR-RFLP, and M13 RAPD-PCR. The analysis revealed that Bacillus subtilis U170B and B. subtilis U146A isolated from iru were the closest relatives of strains belonging to the phylogeny of B. subtilis sensu stricto and were related to other bacilli isolated from different continents that had functional benefits. The two isolated species exhibited resistance to acidic pH (pH 2.0). The survival rates of B. subtilis U170B, B. subtilis U146A, and B. clausii UBBC-07 (commercial probiotic strain) cultured at pH 2.0 for 3 h were 33.45, 12.44, and 9.53%, respectively. The strains were highly tolerant to bile salts [0.3% (w/v)]. B. subtilis U170B exhibited the highest cell viability (43.45%) when cultured for 3 h in the presence of bile salts, followed by B. subtilis U146A (25%) and B. clausii UBBC-07 (18.94%). B. subtilis U170B and B. subtilis U146A did not exhibit haemolytic activity and were susceptible to different antibiotics. Additionally, these two strains exhibited weak antagonistic activity against B. cereus. The diverse wild strains of B. subtilis can be used as a safe multifunctional starter culture for the industrial production of condiments with health benefits.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.1
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• pp.23-30
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• 2014

The present study investigated the effects of dietary Edwardsiella tarda (E. tarda) specific bacteriophage (phage) and Bacillus subtilis (B. subtilis) mixture on innate immune responses and antibacterial activity of Nile tilapia, Oreochromis niloticus. In a dietary experiment, tilapia were fed the control diet (C), a phage-only supplemented diet (P), a B. subtilis only supplemented diet (B), or a B. subtilis and phage mixed diet (B+P). A respiratory burst and significant increase in lysozyme activity (P<0.05) were noted in the B+P group, as compared to other groups after 4 days of feeding. The B group showed a significant (P<0.05) increase in respiratory burst and lysozyme activity versus the C and P groups, whereas no significant increases (P<0.05) were observed in the P and C groups. $ACH_{50}$ was significantly up-regulated in the B+P group versus other groups after 8 days of feeding (P<0.05). In vivo antibacterial activity was significantly enhanced in the B+P fed group, as compared to other groups (P<0.05) after 7 days of E. tarda challenge. A significant (P<0.05) increase in antibacterial activity was seen in the B group, as compared to C or P groups after 14 days of feeding. These results suggest that a B. subtilis and phage mixture could be utilized as an alternative to antibiotics in the control of fish diseases caused by E. tarda.

• Microbiology and Biotechnology Letters
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• v.19 no.4
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• pp.356-361
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• 1991

- To investigate the possibility of genetic development for a multi-purpose strain of Bacillus subtilis YBL-7 against Fusat-iurn solani causing root rot of many impotant corps, the plasmid pGU66 inserting urease gene of Bacillus pasteurii had been introduced into Bacillus subtilis YBL-7 by PEG-induced protoplast (PIP) transformation system. Protoplasts of B. subtilis YBL-7 were prepared by treating the cells with lysozyme (200 $\mu g$/ml) in hypertonic buffer (SMMP). The highest transformation frequency was achieved when cells of the strain with lysozyme at $42^{\circ}C$ for 90 minutes. Optimal transformation was obtained using polyethylene glycol (MW 4000) at final concentration of 30% (V/V). The transformation frequency was increased proportionally to 1.2 $\mu g$ of plasmid DNA. At best condition, the transformation frequency (transformants/ regenerants/$\mu g$ of DNA) for pGU66 was appoximately $4 \times 10^{-3}$. Also, the urease gene was strongly expressed in the transformants of B. subtilis YBL-7 and maintained steadily. The antifungal ability of transformant was very similar to that of B. ssubtilis YBL-7.

• Journal of the Korean Society of Food Science and Nutrition
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• v.15 no.4
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• pp.1-9
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• 1986

The mashes of soybean paste were preparea using the conventional meju fermented naturally by wild microoganisms or the new types of meju fermented by pure cultures of Aspergillus oryzae, Bacillus natto and B. subtilis to elucidate changes during the aging period. The results obtained are as follows ; The soybean paste made with conventional meiu and Asp. oryzae meju showed higher content of amino nitrogen than those of B, natto and B. subtilis meju. Soybean paste made with conventional meju contained a little more content of total and reducing sugars than other soybean pastes. ph during aging period was higher than 5.0 for the Asp. oryzae paste while less than 4.5 for B. subtilis paste. Aspartic acid. threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionne, leucine and histidine for Asp. oryzae paste ; tyrosine, arginine and proline for conventional meju paste; and isoleucine and phenylalanine for B. subtilis paste were found to be peak amount 90 days after the preparation. The content of total free amino acid was high in the order of Asp. oryzae paste, conventional paste, B. natto paste and B. subtilis paste.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.262-273
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• 1987

The replication region of the chloramphenical resistance plasmid pSBK203 of Staphylococcus aureus was cloned using pBR322 and pBD9 as vectors. Cloned replication tegion and chloramphenicol resistance gene were recombined to pBR322. The reconstructed vector behaved as a shuttle vector for E. coli and B. subtilis.

• Journal of Life Science
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• v.14 no.3
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• pp.391-395
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• 2004

To overproduce the cyclodextrin glucanotransferase(CGTase) of Bacillus stearothermophilus NO2 in B. subtilis, the pJH-CGTl plasmid (8.14 kb) was constructed, in which the ORF of CGTase gene could be transcribed by strong constitutive promoter, P$\_$JH/. To overproduce CGTase from a recombinant B. subtilis, the effect of media on the cell growth and expression level of CGTase were investigated with various media (LB, 2${\times}$LB, 5% molasses+2% CSL, CS, LBG) in the flask culture. Among them, [5% molasses+2% CSL] medium revealed the maximum expression level of CGTase with 1.8 unit/$m\ell$ at 9 hr culture. In the batch culture on [10% molasses+5% corn steep liquor] medium the expression level of CGTase, the secretion efficiency, and plasmid stability were about 4.2 unit/$m\ell$, 90% and 90%, respectively, at 30 hr culture. The cell growth and expression level in the fermenter culture with the industrial molasses medium were increased by 2-folds over the flask culture.

• Microbiology and Biotechnology Letters
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• v.30 no.3
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• pp.210-215
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• 2002

A bacterium degrading soy protein was isolated from Korean traditional fermented foods. The isolated strain was identified as Bacillus subtilis, and named as B. subtilis EB464. The optimum pH and temperature of the protease produced by 5. subtilis EB464 were pH 9.0 and $50^{\circ}C$, respectively. The protease was stable in the range of pH 6~10 and below $40^{\circ}C$. The content of water-soluble protein and free amino acid of the medium were increased from 4.2% to 20.6% and ken 1.9% to 22.0%, respectively, by solid-state fermentation of soybean meal with B. subtilis EB464 for 72 h.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.5
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• pp.600-605
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• 2006

B. subtilis JM3 pretense from naturally fermented anchovy sauce was purified in $40{\sim}60%$ ammonium sulfate fraction. In order to accelerate the fermentation of anchovy sauce,2% and 4% of crude B. subtilis JM3 protease were added to 6 month-fermented anchovy sauces, respectively and then the various quality characteristics such as pH, lactic acid, amino-nitrogen, VBN, browning and hydrolysis degree, VBN, and sensory evaluation were analyzed at different storage times. pH was constant during storage time in all samples, whereas lactic acid contents of anchovy sauces hydrolyzed by 2% and 4% proteases were higher than that of control. The amino-nitrogen and volatile basic nitrogen contents of anchovy sauce with 2% and 4% proteases were twice higher than those of control. Anchovy sauces with 2% and 4% pretense increased the hydrolysis rate by 27% and 32%, respectively. Browning degree of anchovy sauce with 4% was higher than those of 2% and control. Anchovy sauce with 2% and 4% proteases was good in sensory evaluation of color, aroma, and taste attributes.