• Food Science and Preservation
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• v.7 no.4
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• pp.414-417
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• 2000

Competent cell transformation of B. subtilis AC819 was carried out using phenotypic protease-defective(Npr-) DNA of B. subtilis MT-2. An obtained transformant, designated B. subtilis HL-1, was obtained by homologous DNA recombination. Phenotypes of B. subtilis HL-1 were characterized histidine requirement streptomycin-resistance, tetracyclin resistance and non-producing protease. Protoplast transformation frequency of B. subtilis HL-1 by plasmid pUB110 was higher than that of B. subtilis MT-2. From this result, B. subtilis HL-1 is useful for protease gene transformation and thermostable protease gene cloning as a host.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1092-1100
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• 2012

Ten Bacillus strains with antimicrobial activities were isolated from Cheonggukjang produced at different parts in Korea. They all inhibited Listeria monocytogenes ATCC 19111 and nine inhibited Bacillus cereus ATCC 14579. Four isolates (W42, H27, SKE 12, and K21) showing strong inhibiting activities were identified as B. subtilis. B. subtilis W42 was the most inhibiting strain. The antimicrobial activity of culture supernatant from B. subtilis W42 was destroyed completely by proteinase K treatment, indicating that a bacteriocin was the responsible agent. The bacteriocin, Bac W42, was most stable at pH 7 and stable between pH 3-6 and 8-9. Bac W42 was stable up to $80^{\circ}C$. BHI (brain heart infusion) and TSB (tryptic soy broth) were the best media for the activity (320 AU/ml) followed by LB (160 AU/ml). Bac W42 was partially purified by column chromatographies. The specific activity was increased from 1,151.2 AU/ml to 9,043.5 AU/ml and the final yield was 26.3%. Bac W42 was 5.4 kDa in size as determined by SDS-PAGE. Bac W42 showed bactericidal activity against L. monocytogenes ATCC 19111.

• Microbiology and Biotechnology Letters
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• v.19 no.6
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• pp.614-618
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• 1991

Antifungal compound, KRF-001, was produced by Bacillus subtilis subsp. krictiensis isolated from soil. Physico-chemical factors affecting cell growth and bioactivity were examined to improve the production yield. Nutrient composition, temperature, pH and phosphate ion concentration were proved to be important factors for the production of KRF-001. Mutation was performed to select high yielding strains. First, mutation was performed with ultra-violet light, and the second mutation process was conducted by MNNG (N-Methyl-N'-nitro-N-nitrosoguanidine) resulting in three high yielding strains.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.677-680
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• 2007

To analyze a cotG-based Bacillus subtilis spore display system directly, $GFP_{UV}$ was expressed on the surface of Bacillus subtilis spores. When $GFP_{UV}$ was fused to the C-terminal of the cotG structural gene and expressed, the existence of a $CotG-GFP_{UV}$ fusion protein on the B. subtilis spore was confirmed by flow cytometry confocal microscopic analysis. When the cotG anchoring motif was deleted, no fluorescence emission was observed under flow cytometry and confocal microscopic analysis from the purified spore, confirming the essential role of CotG as an anchoring motif. This $GFP_{UV}$ displaying spore might be used for another signaling application triggered by intracellular or extracellular stimuli.

• Journal of Ginseng Research
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• v.28 no.1
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• pp.67-70
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• 2004

Bacillus subtilis B-4228 selected from ginseng field soil for prevention of rusty root was tested for the control of ginseng root rot. In petri-plate dual culture, mycelial growth of Cylindrocarpon destructans was inhibited by B-4228 and hyphal swelling of C. destructans was occurred. In pot experiment with C. destructans-contaminated soil B-4228 dipping of ginseng seedling showed significant preventive effect of root rot (p=0.01), percent healthy root 82% and 20% for treatment and control, root rot rate 6% and 50.4%, respectively.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.66-66
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• 2018

Light fermentation has been conducted under different light conditions such as normal dark light, white light, and light emitting diodes (LEDs) various color (blue, green, red, white on blueberry powder with fermenting bacteria Bacillus subtilis (B2). The bacteria B2 was isolated and identified by 16S rRNA sequencing method. RYRP biologically converted to secondary metabolites through light fermentation in the presence of Bacillus subtilis, the bacteria actively involved in bioconversion process. LEDs fermentation to enhance the production of phenolic content while comparing to normal dark and white light. Among the different color LEDs, blue LEDs mediated fermentation showed higher amount of total phenolic and flavonoid content. Then blue LEDs mediated fermented compound were characterized by FTIR and GC-MS, subsequently the compound was analyzed antioxidant activity tests and the antioxidant activity exhibited higher. This is the first study to demonstrate that B. subtilis-LEDs mediated fermentation is useful for facilitating phenolic compound production and enhancing antioxidant activity, which may have greater application fermentation fields.

• Journal of Microbiology and Biotechnology
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• v.21 no.11
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• pp.1193-1198
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• 2011

Encapsulation of biological material in the permiselective membrane allows to construct a system separating cells from their products, which may find biotechnological as well as biomedical applications in biological processes regulation. Application of a permiselective membrane allows avoiding an attack of the implanted microorganisms on the host. Our aim was to evaluate the performance of Bacillus subtilis encapsulated in an elaborate membrane system producing listeriolysin O, a cytolysin from Listeria monocytogenes, with chosen eukaryotic cells for future application in anticancer treatment. The system of encapsulating in membrane live Bacillus subtilis BR1-S secreting listeriolysin O was proven to exert the effective cytotoxic activity on eukaryotic cells. Interestingly, listeriolysin O showed selective cytotoxic activity on eukaryotic cells: more human leukemia Jurkat T cells were killed than human chronic lymphocytic B cells leukemia at similar conditions in vitro. This system of encapsulated B. subtilis, continuously releasing bacterial products, may affect selectively different types of cells and may have future application in local anticancer treatment.

• Mycobiology
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• v.45 no.4
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• pp.385-391
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• 2017

The ability of Bacillus subtilis, strain ALICA to produce three mycolytic enzymes (chitinase, ${\beta}$-1,3-glucanase, and protease), was carried out by the chemical standard methods. Bacillus subtilis ALICA was screened based on their antifungal activity in dual plate assay and cell-free culture filtrate (25%) against five different phytopathogenic fungi Alternaria alternata, Macrophomina sp., Colletotrichum gloeosporioides, Botrytis cinerea, and Sclerotium rolfesii. The B. subtilis ALICA detected positive for chitinase, ${\beta}$-1,3-glucanase and protease enzymes. Fungal growth inhibition by both strain ALICA and its cell-free culture filtrate ranged from 51.36% to 86.3% and 38.43% to 68.6%, respectively. Moreover, hyphal morphological changes like damage, broken, swelling, distortions abnormal morphology were observed. Genes expression of protease, ${\beta}$-1,3-glucanase, and lipopeptides (subtilosin and subtilisin) were confirmed their presence in the supernatant of strain ALICA. Our findings indicated that strain ALICA provided a broad spectrum of antifungal activities against various phytopathogenic fungi and may be a potential effective alternative to chemical fungicides.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.274-281
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• 2006

A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.

• Journal of fish pathology
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• v.26 no.3
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• pp.185-191
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• 2013

The present study was performed to investigate an antibacterial activity of specific bacteriophage (phage) and Bacillus subtilis KM-1 (B. subtilis) mixture against Edwardsiella tarda (E. tarda). An appropriate number of phage showing the most effective antibacterial activity was $2{\times}10^5$ PFU/ml with $1{\times}10^7$ CFU/ml of B. subtilis 36 h post incubation. On the other hand, B. subtilis showed a dose dependant manner in inducing antibacterial activity in the presence of phage ($2{\times}10^5$ PFU/ml). The phage and B. subtilis mixture showed higher antibacterial activity against E. tarda than phage or B. subtilis only. These results suggest that the phage and B. subtilis mixture could be utilized as an alternative to antibiotics in the control of fish diseases caused by E. tarda.