• Journal of Environmental Health Sciences
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• v.31 no.5 s.86
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• pp.444-450
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• 2005

The present study was designed to investigate whether traditional Korean fermented soybean paste(doen-jang) and soybean extracts have inhibitory effects on the growth of KB cell, an oral epithelioid cancer cell. When KB cell ATCC CCL-17 was cultivated for 48 flours with the addition of 0.5% of the five types or doen-jang extract, the growth of KB cell was inhibited by all types of extract, and ethyl acetate extract showed the highest inhibitory effect. In case of soybean extract, all types of extract also showed KB cell inhibitory effects, however, generally less than those of doen-jang extract. When ethyl acetate extract of doen-jang was added in different concentrations and KB cell was cultivated for 24 hours and 45 hours, strong inhibitory effect began to appear from the concentration of 1.25 mg/ml. Although soybean extract showed such a tendency, its effect was lower than that of doen-jang extract. These results indicate that doen-jang extract has inhibitory effect against KB cell, and particularly ethyl acetate extract has the highest effect. The effect of doen-jang extract might be possibly enhanced by the fermentation of soybeans. It is assumed that doen-jang extract may be used to develop nontoxic medicines for preventing and treating oral diseases.

• The Korean Journal of Food And Nutrition
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• v.16 no.3
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• pp.218-223
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• 2003

This study is proceeded on the development of standard method for making soybean paste by Korean traditional method. Fermentation condition of Meju was 1) Pre-fermentation : 30 days in about 20$^{\circ}C$ room, 2) Main fermentation : 5 days in 30$^{\circ}C$ or upper temperature, 3) Post fermentation and drying : 30 days in well sunlightened room in January. Meju was soaked in 18% salt solution(Meju 7kg/salt solution 20L) for 35～40 days (from late February to early April) and after soaking Meju was filtered as unsoluble solute and crushed and put into traditional Korean receptacle(named 'Dok'). Crushed Meju was stored from early April to mid September and Meju was changed into soybean paste(Doen-jang). During fermentation amino acid nitrogen in Doen-jang was slightly increased in early period and decreased lately. It has been proved that by panel test soybean paste made by the method suggested in this study was more excellent than commercially fermented soybean paste. This study has presented the possibility of commercial production of soybean paste made by traditional method.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.87-90
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• 1998

Various bacterial strains that secret extracellular fibrinolytic enzyme were screened from Doen-Jang, a traditional soybean fermented food in Korea. Five microbes of them were identified to be Bacillus sp. strains according to Bergey's manual of systematic bacteriology. The culture filtrates of B. amyloliquefaciens (2.46 plasmin unit/ml) and B. pantothenticus (3.82 plasmin unit/ml) showed a level of fibrinolytic activity that was about three times higher than that of plasmin 1.0 unit and Bacillus subtilis showed the highest fibrinolytic activity (4.94 plasmin unit/ml). All of the extracellular proteases showing the fibrinolytic activity are confirmed by SDS-PAGE followed by reverse fibrin zymogram activity assay and we proposed that some of the fibrinolytic enzymes from this work are novel enzymes.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.72-79
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• 2005

A new Bacillus peptidase, bacillopeptidase DJ-2 (bpDJ-2), with molecular mass of 42 kDa and isoelectric point (pI) of 3.5- 3.7, was purified to homogeneity from Bacillus sp. DJ-2 isolated from Doen-Jang, a traditional Korean soybean fermented food. The enzyme was identified as an extracellular serine fibrinolytic protease. The optimal conditions for the reaction were pH 9.0 and $60^{\circ}C$. The first 18 amino acid residues of the N-terminal amino acid sequence of bpDJ-2 were TDGVEWNVDQIDAPKAW, which is identical to that of bacillopeptidase F (bpf). However, based on their Nterminal amino acid sequence, molecular size, and pI, it is different from that of bpf and extracellular 90 kDa. The whole (2,541 bp, full-bpDJ-2) and mature (1,956 bp, mature-bpDJ-2) genes were cloned, and its nucleotide sequence and deduced amino acid sequence were determined. The expressed proteins, full-bpDJ-2 and mature-bpDJ-2, were detected on SDSPAGE at expected sizes of 92 and 68 kDa, respectively.

• Journal of Environmental Health Sciences
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• v.34 no.3
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• pp.207-212
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• 2008

This study was performed to investigate the anti-microbial activity of extract from Korean fermented soybean paste (doen-jang) against 16 types of oral microbes, and to determine the minimum inhibition concentration (MIC) of the extract for three major microbes causing human oral diseases (Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans). The extract was prepared using ethyl acetate and it was treated with the oral microbes at a concentration of 5.00 mg/ml (0.5%). The anti-microbial activity and MIC were measured using broth dilution method. Significant reduction of microbial activities of 16 types of oral microbes occurred when the soybean paste extract was added to the broth compared to the control (p<0.01), and striking inhibition (more than 99%) was observed in ten types. S. mutans, which causes dental caries, showed MIC at a concentration of 1.25 mg/ml for the extract. P. gingivalis, which causes adult periodontal disease, showed MIC at a concentration of 2.50 mg/ml for the extract. C. albicans, which causes denture stomatitis and angular stomatitis, showed MIC at a concentration of 20 mg/ml for the extract. These results indicate that ethyl acetate extract of doen-jang showed strong anti-microbial effect against 16 types of oral microbes, and the anti-microbial effect of the extract against oral microbes was stronger against bacteria than against fungi. The anti-microbial effect might be possibly enhanced by the fermentation of soybeans.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.649-656
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• 2002

Fibrinolytic microorganisms were screened from 42 samples of Korean fermented food (7 kinds of Chungook-jang, 14 kinds of commercial Doen-Jang, 5 kinds of home-made Doen-jang, and 16 kinds of Jeot-gal), 15 samples of Japanese fermented food (5 kinds of home-made soybean paste, and 10 kinds of Natto), and 19 samples of Indonesian fermented food (Tempe) as well as starters of Meju (500 microflora from Korea, and 22 from China). Initially, 11 isolates with strong fibrinolytic activity were selected for further characterization. The fibrinolytic activity of the 11 isolates ranged from 89 to 199% of standard plasmin. Four strains, M5l from Korean fermented food (Meju), I 1-1, I 1-4, and I 5-1 from Indonesian fermented food (Tempe), were chosen based on the degree of activity and reproducibility, and identified as Staphylococcus sciuri, Citrobacter or Enterobacter, Enterococcus faecalis, and Bacillus subtilis, respectively. The first two isolates are pathogenic stains while the latter two are considered as GRAS (Generally Recognized As Safe). Fibrinolytic activity of E. faecalis, characterized and designated as BRCA-5, reached a maximum, when the producer was cultivated in Ml7 broth supplemented with 1.0% glucose for 5 h at 37$^{\circ}C$ with shaking at 180 rpm. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of 5. faecaiis BRCA-5 showed stronger activity than plasmin and streptokinase, but similar degree of specific activity as nattokinase and urokinase, aud it also demonstrated anticoagulant and antiplatelet activity ex vivo. These features of E. faecalis make it an attractive agent as a biomaterial for health-promoting foods.

• Proceedings of the Korean Environmental Health Society Conference
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• 2005.11a
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• pp.101-103
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• 2005

This study was performed to investigate the changes of Bacillus sp. of traditional Korean soy sauce (kan-jang) and soybean paste (doen-jang) during the ripening and storage for 12 months. All of the preparation methods for soy sauce and soybean paste followed the recommendations of the Korea Food Research Institute. The soy sauce and soybean paste were analyzed at 0, 6, and 12 months. The numbers of Bacillus sp. of meju (soybean cakes) or soy sauce and soybean paste at the initial stage were significantly higher than those of soybeans (p<0.05). The number of Bacillus sp. increased in soy sauce and soybean paste after 12 months of ripening and storage. This study indicates that this comes from the preparation and fermentation of meju. It is suspected that the organoleptic characteristics of soy sauce and soybean paste might be inferior over 1 year of storage time. Therefore, more research fur overall microbiological quality changes of soy sauce and soybean paste during the storage period should be conducted to interpretate this characteristics more accurately.

• Proceedings of the Korean Environmental Health Society Conference
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• 2004.06a
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• pp.182-184
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• 2004

Oral cancer is the sixth most common cancer globally. The effects of several extracts from soybeans and Korean soybean paste (doen-jang) on the growth of human oral carcinoma cells in vitro were assessed. We prepared petroleum ether extract, ethyl acetate extract, chloroform extract, methanol extract, and water extract from soybeans and soybean paste. We used KB cell, which is an oral epidermoid carcinoma cell, and investigated proliferation of the tumor cells using MTT method. Each extract of soybean paste suppressed the KB cell proliferation. A dose-response relationship was observed between the level of ethyl acetate extract of soybean paste and its suppression of the cell proliferation. The effects of soybean extracts were lower than those of soybean paste extracts. The effects might be enhanced by the fermentation of soybeans. The results of this work indicate that extracts from soybeans and Korean soybean paste could have potential as anti-tumor substances.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1111-1114
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• 2001

An extracellular fibrinolytic-enzyme-producing bacterium was isolated from Doen-Jang, a Korean traditional fermented flood, and identified as Bacillus sp. DJ based on its morphology and cellular fatty acid composition. The total extracellular fibrinase (EF) from Bacillus sp. DJ was analyzed using three fibrin zymographic techniques, SDS-fibrin zymography (SDS-FZ), isoelectrofocucing-fibrin zymographs(IEF-FZ), and a two-dimensional SDS-fibrin zymographic analysis (2D SDS-FZ). As a result, the EP map of Bacillus sp. DJ was established. The results suggest that the 2D SDS-FZ method will be a useful tool for the proteomic approach for many other bacterial pretenses.

• BMB Reports
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• v.34 no.2
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• pp.134-138
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• 2001

By adding sodium chloride (2.5%) into a Bacillus amyloliquefaciens DJ-4 culture broth, two serine-type fibrinolytic proteases with a molecular weight of 29 (subtilisin DJ-4) and 38-kDa were stimulated on the SDS-fibrin zymogram or inhibitor gels. B. amyloliquefaciens DJ-4 showed the highest proteolytic activity (5.52 plasmin NIH unit/ml) on the fibrin plate based on the molar ratio when cells were subjected to the 2.5% NaCl. Using a fibrin plate, the secreted protease from this strain in the presence of 5% NaCl showed that about 49% of the enzyme's activity remained after incubation at $60^{\circ}C$ for 30 min, but as the salt concentration was increased (10% NaCl) the activity nearly disappeared (0.14 plasmin NIH unit/ml). However, through a fibrin zymography assay, three fibrinolytic enzymes (38, 53 and 80-kDa) from the cells in the presence of 10% NaCl were detected. Also, two salt-activated serine-type fibrinolytic professes (29 and 38kDa) showed thermostability from 65 to $70^{\circ}C$ for 30 min. Furthermore, these professes also showed stability, pH 6-11. In particular, 29-kDa (subtilisin DJ-4) was very stable in the pH range of 4-11 at $4^{\circ}C$ for 48 h.